▪ Position of hand Downward

▪ Handwashing song Happy birthday song (twice)

GENERAL LABORATORY 12) HEPA (high efficiency particulate air) filter can remove particles of at least: 0.3
microns in size
LABORARTORY SAFETY
1) Safety diamond SPECIMEN COLLECTION
Health (LEFT) = BLUE Flammability/Fire (TOP) = RED 1) BASAL STATE is usually taken early in the morning, before the patient has eaten or
Stability/Reactivity (RIGHT) = YELLOW Special/Specific info (BOTTOM) = WHITE before patient becomes physically active
2) Magnitude of Hazard (Health) 2) Number of fasting hours is part of: PATIENT PREPARATION
0 – none 1 – slight 2 – moderate 3 – serious e treme 3) Puncture site is disinfected/cleansed using: ISOPROPYL ALCOHOL
3) Waste Segregation 4) Antiseptic used in ethanol testing BENZALKONIUM CHLORIDE
Dry, non-infectious = BLACK Sharps = RED 5) Photosensitive analytes: BILIRUBIN, CK
Wet, non-infectious = GREEN Radioactive wastes = ORANGE 6) Analytes that require chilling of specimen with ice: AMMONIA, BLOOD GAS
Wet, infectious = YELLOW 7) ANALYTES WITH DIURNAL VARIATION
4) Types of Fire INCREASED IN AM = ACTH, cortisol, iron, Aldosterone
Paper, cloth, wood = CLASS A Electrical equipment = CLASS C INCREASED IN PM = ACP, GH, PTH, TSH
Flammable liquids = CLASS B Combustible metals = CLASS D
8) Sources of error in ammonia analysis: SMOKING AND DETERGENT
Arsenal fire (liable to detonation) = Class E 9) ANALYTES AFFECTED BY TOBACCO SMOKING
5) Steps to do when fire is discovered INCREASED: plasma catecholamines, serum cortisol, plasma fatty free acids
R = RESCUE A = ALARM C = CONTAIN E = EXTINGUISH lactate, insulin, epinephrine, growth hormone, 5-HIAA (in urine)
6) Steps in using fire extinguisher leukocyte (WBC) count; neutrophils; monocytes
Pull the pin; Aim at the base of fire; Squeeze handle; Sweeping motion blood carboxyhemoglobin levels, erythrocyte (RBC) count,
Types of Fire Extinguisher Hb concentration, Hct, MCV, IgE
Extinguishers that contain halon can be used for fire class(es): BC DECREASED: IgA, IgG, and IgM; Eosinophils; Vitamin B12
Extinguisher that contains dry chemical can be used for fire class(es): ABC decreased sperm counts and motility
When skin contact with chemical occurs, the best first aid is to f lush the area with 10) ANALYTES INCREASED IN ALCOHOLISM
large amounts of water for at least 15 minutes and seek for medical attention. = Aminotransferases, lipoproteins, bilirubin, ketone bodies, triglycerides
Hazard associated with breakage in centrifuge = AEROSOL/ N HAZARD ANALYTES DECREASED IN ALCOHOLISM
Components of Chain of Infection (6 components) = Glucose, albumin, transferrin
Source of infection (Pathogen), Reservoir, Portal of Exit, Mode of 11) Effect of bilirubin and hemoglobin in ALP determinations: INCREASED ALP
Transmission, Portal of Entry, Susceptible Host 12) Analyte detects to assess completeness of a 24-hour urine: CREATININE
11) Single most effective way of preventing transmission of disease: HANDWASHING 13) Specimen used in Newborn Screening: CAPILLARY BLOOD (BLOOD SPOT)
▪ Steps in Handwashing: Wet, Lather, Scrub, Rinse, Dry 14) Preferred sample for electrolyte analysis: HEPARINIZED PLASMA
▪ Length of Scrubbing 20 seconds (CDC) 15) Preferred specimen for ammonia analysis: EDTA PLASMA/HEPARINIZED PLASMA

1
 Vacutainer tubes that contains EDTA: LAVENDER, PINK, WHITE, TAN g. A positive displacement pipet operates by moving the piston in the pipet tip or
Specimen for ACP determination: SERUM, VAGINAL WASHING barrel, much like a hypodermic needle. It does not require a different tip for
Effect of using non-anticoagulated tube for ACP: NONE each use. Because of carry over concerns, rinsing and blotting between samples
Effect of direct rectal exam in ACP levels: INCREASED may be required.
Effect of intramuscular injection in CK-MM levels: INCREASED Analyzer characterized by continuous system of tubing: CONTINUOUS FLOW
Effect of breast examination on prolactin levels: INCREASED ANALYZER
Bilirubin Level of Icteric sample: . Primary source of error in continuous flow analyzer:
Bilirubin Level associated with jaundice: m /d . Reactants in the continuous tubing are separated using:
Kernicterus – accumulation of bilirubin in the brain; danger of kernicterus is at c. Purpose of coiled tubings in continuous flow analyzer M N
certainty at levels exceeding 20mg/dL d. Example of Continuous Flow Analyzer: AUTOANALYZER, TECHNICON
Bilirubin levels of 25mg/L (430mmol/L) can cause interference in albumin (in Analyzer characterized by spinning rotor: CENTRIFUGAL ANALYZER
HABA method), in cholesterol (in assays using ferric chloride reagent) and in total Most popular type of analyzer; analyzer type that can perform random access,
protein assays (using Biuret reaction) batch and sequential analysis: DISCRETE ANALYZER
Fasting requirement for Lipid analysis: AT LEAST 12 HOURS Examples of Discrete Analyzers
Lipemia occurs when TAG level exceeds 400mg/dL. It causes interference for ACA (Dupont) ASTRA/SYNCHRON (Beckman)
amylase, urates, urea, creatine kinase, bilirubin and total protein CENTRIFICHEM (Baker) COULTER CHEMISTRY (Coulter Electronics)
Blood volume of an average person: KDA (American Monitor) ARCHITECT and TDX (Abbott)
males: 5-7L females: 4-6L PARAMAX HITACHI (Boehringer Mannheim/Roche)
29) Preferred vein for venipuncture: VITROS (Ortho-Clinical D.) ADVIA and DIMENSION (Siemens)
30) Analytical Testing performed outside the laboratory is known as: NANOMETER is the unit of measure for the wavelength of radiant energy (light).
Wavelength is inversely proportional to the amount of energy. The longer the
INSTRUMENTATION AND AUTOMATION wavelength, the lower the energy; the shorter the wavelength, the higher the
Centrifuge energy.
. Centrifuge type recommended for Serum Separator Tubes (SST) N Spectra of Light
C N / N N C C N
. Visible Light Spectrum nm
. Disinfection of centrifuge is done :
. Spectrum of UV light nm nm
c. Calibration for speed of centrifuge is done every: M N
c. Spectrum of IR light 7 0nm – 0.3cm
d. Device used to calibrate the speed of centrifuge: C M /
Color and Energy
Pipets
. Color with the highest energy VIOLET
. Pipet with the highest precision and accuracy: M C
b. Color with the longest wavelength RED
. Transfer pipet used for viscous solutions: N
11) BEER-LAMBER LAW = states that absorbance (optical density) is directly
c. Pipets with etched rings or continuous rings:
proportional to concentration
d. Pipets without graduations lines at the tip: M
A = 2 – log%T A = abc
e. Cleaning solution for glassware: C C M
. Sterilization technique for glassware: N C N

2
 Graph of Beer’s Law will show a straight line if:
Concentration vs Absorbance are plotted using = LINEAR PAPER
Concentration vs r s i ce are plotted using = SEMILOG PAPER
Used to correct for interferences attributed from the sample: SAMPLE BLANK
Part of the spectrophotometer that isolates the specific wavelength of interest
(desired wavelength): MONOCHROMATOR (Prisms, Diffraction Gratings,
Interference filters, Colored glass filters)
Part of spectrophotometer that converts light to electrical energy:
PHOTODETECTOR
Instrument with 2 photodetectors
DOUBLE BEAM IN SPACE SPECTROPHOTOMETRY
17) Instrument with 2 monochromators, positioned at 90deg: FLUOROMETRY
a. Characteristics of excitation light: HIGH ENERGY, SHORT WAVELENGTH
b. Characteristics of fluorescent light: LOW ENERGY, LONGER WAVELENGTH
c. Disadvantage of Fluorometry: QUENCHING
d. Effect of Quenching in fluorescence: DECREASED FLUORESCENCE
18) Instrument that measures the light/color emitted by the analyte; used for analytes
that are easily excited by the flame = FLAME EMISSION PHOTOMETRY
a. Color emitted by Na = YELLOW K = VIOLET Li = RED
b. Dilution of serum used in Na/K analysis in FEP = 1:100 or 1:200
c. Internal standards used in FEP = LITHIUM, CESIUM
19) Instrument used for analytes that are NOT EASILY excited by the flame; serves as
reference method for CALCIUM and MAGNESIUM = ATOMIC ABSORPTION
SPECTROPHOTOMETRY
a. Light source of AAS = HOLLOW CATHODE LAMP
b. Purpose of the flame in AAS = ATOMIZER
c. After analyzing an analyte, the hollow cathode lamp must be changed first,
before proceeding with the analysis of the next analyte
20) Instrument that measures reflected light; serves as the principle behind Vitros,
Kodak Ektachem and Automated Dipstick Readers: REFLECTANCE
SPECTROPHOTOMETRY
Dry Slide Technology (layers; top to bottom)
Spreader → Scavenger → Reagent → Indicator → Support/Basal
21) Instrument that measures light scattered: NEPHELOMETRY
Instrument that measures light blocked by particles: TURBIDIMETRY
22) ELECTROPHORESIS – method that separates substances according to their charge
a. Support media include:
paper, cellulose acetate, agarose gel, polyacrylamide gel, starch gel
b. Positive pole/electrode = ANODE
c. Negative pole/electrode = CATHODE
d. Most common pH used in protein electrophoresis = 8.6
e. In alkaline pH, proteins become = NEGATIVELY CHARGED
f. pH at which substances will have net zero charge
= ISOELECTRIC POINT
Electrodes
. Principle used in O2 determination, glucose id se peroxidase = M M
. Principle used in pH and pCO2 determination = N M
c. Electrode used in pH determination = C
d. Name of electrode used in pO2 determination = Clar e electode
e. Name of electrode used in pCO2 determination = e erin aus electode
. Electrode used in potassium determination = N M CNC
. Parameter that can be used as quality control for ISE = N N
24) Hardware = tangible/physical component of computers; i.e. CPU, keyboard,
mouse, printer, scanner, monitor
Software = non-tangible component of computers; i.e. applications, programs,
operating systems
25) Input = used to enter information into the computer; i.e. keyboard, scanner
Output = used to produce a result from the computer; i.e. monitor, printer
26) First step to be taken when attempting to trouble-shoot electronic equipment is to
ensure that the instrument has been turned off. This is done in order to prevent
electric hazard.
27) In some electrical instruments, there are three wires. The third wire is for
grounding. It reduces the risk of serious electric shock
Quality Assurance
1) Ability to detect small amounts of analyte = N
Ability to detect only the analyte of interest = C C
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2) Closeness of the value to the target/true value = CC C 16) Warning or Screening Rule for Rejection 12S
Closeness of the value to the repeated value = C N 17) Westgard multirule for random error 12S, 13S, R4S
iiy i i ccur cy d recisi 18) Westgard multirule for systematic error 22S, 41S, 10x
3) Indicator of precision = standard deviation 19) Comparison of previous results with the current result DELTA CHECK
Relative indicator of precision = coefficient of variation 20) The next step in the connectivity process is transmittal of test results from the
4) Lower coefficient of variation indicates: GOOD PRECISION data manager to the LIS and/or hospital information system (HIS). This is the
5) Measures the statistical differences between 2 means = t-test INTERFACE PORTION of connectivity. (6th ed Bishop)
Measures the statistical differences between 2 SDs or 2 variances = F-test 21) ASAP means “as soon as possible” and stat is an American medical term
6) Gaussian curve = bell-shaped curve; assumes that mean = median = mode meaning “immediately” (from the Latin “statim”).
7) Bull’s eye in the quality control chart indicates: = CC C
C N Reagent Preparation and Laboratory Mathematics
8) Type of QC where analysis of control samples together with INTRALAB/ 1) Conversion Factors:
patient specimen INTERNAL QC ▪ Bilirubin . ▪ Glucose .
9) Control are materials of known value that is analyzed with patient samples to ▪ Albumin ▪ Thyroxine (T4) .
determine acceptability of results. It is of known quality with several analytes ▪ Na /K+ + ▪ Total protein
present. It resembles patient serum. Controls are usually made from pooled serum. ▪ TAG . ▪ Cholesterol .
▪ Uric Acid . ▪ BUN .
10) Variation/Error present in all measurements, due to chance RANDOM ERROR
11) Causes of random error = mislabeling of sample, pipetting errors, improper 2) SI units used for:
mixing of sample and reagent, voltage fluctuations, temperature fluctuations ▪ Glucose ▪ Creatinine u
Causes of systematic error = improper calibration, deterioration of reagents, ▪ Chloride ▪ Bilirubin u
sample instability, changes in standard materials, instrument drift ▪ CO2 ▪ O2
12) Levey-Jennings Chart (Shewhart Plot) – most widely used quality control chart 3) Panic/Alert/Critical Values
X-axis = independent variable; horizontal line; date or time Glucose d d Bilirubin d
Y-axis = dependent variable; vertical line; analyte concentration 4) Anion Gap = H 5) Carbonic acid = .
13) Main cause of trend = deterioration of reagents 6) Free T4 index = total T4 X THBR
Main cause of shift = improper calibration of instrument 7) g of NaCl combined with 100mL of water to make NSS: 0.85g per 100mL
14) Control values that are far from the main set of values = 8) Amount of NaOCl needed to make 10L of disinfectant: 1L
Westgard Multirule 9) Amount of NaOCl needed to make 7L of disinfectant: 700mL
▪ One control value exceeds 2SD = s
10) What is the concentration in %w/v of a 20g NaOH 4%
▪ One control value exceeds 3SD = s
dissolved in 500mL of water? = (20/500) x 100
▪ The range of 2 consecutive control values are 4 SDs apart = s
▪ Four consecutive control values exceed 1SD = s
▪ Two consecutive control values exceed 2SD = s
▪ Ten consecutive control values are on the same side of the mean =

4
 Carbohydrates
Clinical Chemistry 1) Monosaccharides (Hexoses)
Glucose (dextrose), fructose (levulose, fruit sugar), galactose
Metabolic Blood Test and Analyte detected (Overview) 2) Disaccharides Lactose (milk sugar) = galactose + glucose
Sucrose (table sugar) = fructose + glucose
▪ Nelson-Somogyi method uc se
Maltose = glucose + glucose
▪ Dubowski reaction uc se
3) Hormones:
▪ Van Handel Zilversmit method ri ycerides
a. Decreases blood glucose: insulin
▪ Lieberman Burchard reaction c es er b. Increased blood glucose: glucagon, cortisol, growth hormone, epinephrine,
▪ Biuret Reaction r ei thyroid hormones
▪ Parfentjev reaction i ri e 4) Specimen Collection and Considerations
▪ Jaffe reaction cre i i e a. Standard clinical specimen is fasting venous plasma.
▪ Fearon reaction ure b. Venous blood has lower glucose levels compared to arterial blood
▪ Berthelot reaction ure c. Whole blood gives approximately 10 to 15% LOWER glucose levels than
▪ Caraway method uric cid serum or plasma.
▪ Conway method i d. At room temperature, glucose decreases at a rate of 7 mg/dL/hr
e. At 4°C, glucose decreases by approximately 2 mg/dL/hr
▪ Wacker method H r rd
f. 2 mg of sodium fluoride per mL of whole blood acts as preservative with
▪ Wrobleuski La due H re erse
another anticoagulant (i.e. K+ oxalate); prevents glycolysis for 48-72 hours
▪ Tanzer-Gilvarg r rd
g. If NaF is used alone as an anticoagulant, 3 to 5 times greater concentrations
▪ Oliver-Rosalki or Rosalki-Hess re erse than usual is required (approx. 6 to 10mg of NaF/mL of blood).
▪ Bowers-Comb method h. Fluoride binds magnesium, which causes inhibition of the enzyme enolase.
▪ Bessey-Lowry-Brock i. CSF glucose concentration is approximately 60 to 70% that of plasma
▪ Szasz method concentration.
▪ Ellman method i es er se j. Blood glucose should be obtained 1-2 hours before the spinal tap
▪ Dixon & Purdon, Campbell, Belfield & Goldberg method uc e id se k. 10% contamination with 5% dextrose will increase glucose by 500mg/dL or
▪ Karmen method more
▪ Reitman-Frankel method d In oral glucose tolerance test:
. Blood glucose should return to normal after 2 hours
▪ Schales-Schales ride
. Patient is asked to consume a normal to high carbohydrate intake
▪ Clark-Collip ciu
e s 150g carbohydrates per day for 3 days prior to the test
▪ Titan Yellow method esiu c. Patient should fast at least 8-10 hours
▪ Fiske-Subbarow s e d. OGT testing should be performed on the morning
▪ Kober reaction es r e s e. Blood glucose is taken every 30 minutes for 2 hours.
▪ Porter-Silber reaction c r ic s er ids . Patient should be ambulatory. Glucose load for an adult is 75g.
▪ Zimmerman method e s er ids . Patient should finish drinking the glucose load within 5 minutes
. If patient vomits, discontinue the test.

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6) Long term monitoring (2-3 months) of glucose control = Glycosylated Lipids
Hemoglobin 1) Normal Lipoproteins
a. 1% change is HbA1c is equivalent to m /d ▪ LOW DENSITY LIPOPROTEINS
b. Specimen for HbA1c ole blood - Transports cholesterol from liver to cells; Bad cholesterol
c. HbA1c in patients with hemolytic anemia alsely decreased - Direct relationship to the risk of atherosclerosis and coronary heart
d. Method that is NOT affected by other hemoglobins and temperature disease; 45-50% cholesterol ester by weight
N C M - Beta-lipoprotein; Density: 1.006-1.063
7) Diabetes Mellitus: Type I vs Type II ▪ HIGH DENSITY LIPOPROTEINS
TYPE I DIABETES TYPE II DIABETES - Transports cholesterol from cell to liver;
Frequency 5-10% 90-95% - Reverse cholesterol transport; Good cholesterol
Pathogenesis Def. of insulin Def. of insulin receptor - Inverse relationship to the risk of atherosclerosis and coronary heart
Autoimmune Lifestyle related disease
- Alpha-lipoprotein; 45-55% protein by weight, Density: 1.006-1.21
Previous names Insulin-dependent DM Non-insulin dependent DM
▪ CHYLOMICRONS
Juvenile-onset DM Adult-onset DM
- Lightest (least density) and largest among the lipoproteins
Ketoacidosis Prone to ketoacidosis Not prone to ketoacidosis - 80-95% triglycerides by weight; Density = <0.96
- Transports exogenous triglycerides
8) Glucose intolerance occurring during pregnancy is known as: = N M - Causes turbid or milky appearance of serum after fat ingestion
9) Women who had gestational diabetes are most likely to develop diabetes after ▪ VERY LOW DENSITY LIPOPROTEINS
years - Transports endogenous triglycerides; 45-60% TAG by weight
10) enal t res old for glucose is the level in the blood, above which, glucose is - Pre-beta lipoprotein; Density: <1.006
excreted in the urine. Renal threshold value for glucose is usu y 160-180mg/dL. 2) Reference method for lipoprotein analysis ULTRACENTRIFUGATION
Unit of Sedimentation Rate SVEDBERG UNIT
11) Used to monitor insulin shock andom blood su ar
3) Apolipoproteins: Apo-A1 =
12) Whipple’s triad is associated with: HYPOGLYCEMIA Apo-B48 = C ylomicrons
13) Most common glycogen storage disorder, deficiency VON GIERKE DISEASE Apo-B100 = and
of glucose-6-phosphatase 4) Standing plasma test is used to detect:
Methods ▪ Floating cream layer after overnight refrigeration =
. Non-enzymatic (chemical) glucose methods: Nelson-Somogyi, Folin-Wu, ▪ Floating cream layer + turbid plasma after overnight refrigeration =
Ortho-toluidine 5) Fredrickson’s Classification of Dyslipoproteinemias
. Reagent for Nelson Somogyi = N M ▪ Type I – elevated TAG; presence of chylomicrons
c. Reagent for Folin Wu = M ▪ Type IIa – elevated LDL
d. Positive result in Condensation (o-toluidine) method ▪ Type IIb – elevated LDL and VLDL
= N nm ▪ Type III – elevated cholesterol and TAG; presence of beta-VLDL
e. Reference method for glucose = N ▪ Type IV – elevated VLDL
▪ Type V – elevated VLDL; presence of chylomicrons
. Method affected by reducing and oxidizing agents = C
6) Atherosclerotic plugs/plaques are composed of cholesterol
. Glucose oxidase detects for C

6
7) Least reliable indicator of Coronary Heart Disease (CHD): TRIGLYCERIDES, since 13) LCAT is the enzyme that enhances esterification of cholesterol. It transfers acyl
they have the highest total error, bias, and coefficient of variation groups (fatty acids/esters) in cholesterol.
8) National Cholesterol Program – Guidelines for Acceptable Measurement Error 14) Color Reagents in the Van Handel Zilversmit = chromotrophic acid + H2SO4
(Coefficient of Variations) 15) Form of cholesterol that reacts with cholesterol oxidase = FREE CHOLESTEROL
▪ Total Cholesterol ≤3% Triglycerides ≤5% 16) Cholesterol esterase (cholesterol ester hydrolase) catalyzes the hydrolysis of
▪ HDL-Cholesterol ≤4% LDL-Cholesterol ≤4% cholesterol esters into free cholesterol and fatty acids. This is the first enzyme
9) Positive Risk Factors for Coronary heart disease (NCEP) used in the enzymatic method of cholesterol determination.
Females = 55y/o Males = 45 y/o 17) Lipid stains used in lipoprotein electrophoresis
10) Reference Ranges: National Cholesterol Education Program (ATP III Oil red O, Fat Red 7B and Sudan Black B
Classification) 18) Methods for cholesterol
▪ Total Cholesterol METHODS PROCEDURE
Desirable: <200 mg/dL High: ≥240 mg/dL Zlatkis, Zak & Boyle One-step (Colorimetry)
Borderline high: 200-239 mg/dL iebermann urc ard
▪ LDL-Cholesterol Carr & Drekter Two-step (Extraction & Colorimetry)
Optimal: <100 mg/dL Near Optimal: 100-129 mg/dL Abell Three-step (Extraction, Saponification, Colorimetry)
Borderline High: 130-159 mg/dL High: 160-189 mg/dL Schoenheimer/ Sperry Four-step
Very High: ≥190 mg/dL (Extraction, Saponification, Purification, Colorimetry)
▪ HDL-Cholesterol
Protective against heart disease (Negative Risk for CHD): ≥60
The higher the better: 40-59 Proteins and Liver Function Tests
Major risk factor for heart disease: <40 1) Protein Electrophoresis
▪ TAG Albumin --
Normal: <150 mg/dL High: 200-499 mg/dL Alpha-1 Alpha-1-antitrypsin, alpha-1-acid glycoprotein (orosomucoid), alpha-
Borderline high: 150-199 mg/dL Very High: ≥500 mg/dL fetoprotein, HDL, alpha-1-antichymotrypsin, inter-alpha-trypsin inhibitor,
11) Calculations Gc-globulin
Alpha-2 Alpha-2-macroglobulin, Ceruloplasmin, Haptoglobin
▪ Friedewald Equation
Beta rans errin siderop ilin , Fibrinogen, Complement, C-reactive
LDL in mg/dL = Total Chole – HDL – (TAG/5)
protein, VLDL, LDL, beta-2-microglobulin,
LDL in mmol/L = Total Chole – HDL – (TAG/2.175) Gamma u u i s IgG, IgM, IgA, IgD, IgE
Note: VLDL in mg/dL = TAG/5 or VLDL in mmol/L = TAG/2.175 Proteins and their functions
VLDL is not included in the computation of the LDL. . Indicator of Malnutrition = PREALBUMIN (TRANSTHYRETIN)
▪ Body Mass Index (BMI) . Condition associated with increased albumin = N
below 18.5 = underweight 18.5 to 24.9 = healthy c. Carrier of hemoglobin = HAPTOGLOBIN (haptoglobin is in patients with
25 to 29.9 = overweight 30 or higher = obese hemolytic anemia
12) Abnormal Lipoproteins d. Carrier of iron (Fe3+) = TRANSFERRIN
▪ Beta-VLDL = Floating beta lipoprotein; Type III Dysbetalipoproteinemia e. Carrier of copper = CERULOPLASMIN
▪ Lp(a) = Sinking pre-beta lipoprotein; premature atherosclerosis Decreased ceruloplasmin is associated: N
▪ LpX = Obstructive biliary disease

7
 f. Significance of B-type (brain) natriuretic peptide = C N 9) Bilirubin value taken:
3) Electrophoretic patterns: ▪ before the addition of dissociating/accelerator reagent
a. Beta-gamma bridging C ▪ after the addition of dissociating/accelerator reagent otal bilirubin
b. Monoclonal spike in gamma region M M M 10) elta bilirubin = Bilirubin covalently bound to protein; contributes to direct
c. Increased alpha-2 N C N M bilirubin values
d. Increased alpha-1 and alpha 2 C N MM N ilirubin c e used ssess e cy e i e duc .
e. Decreased alpha-1 = H B1 and B2 levels
4) Cryoglobulins are immunoglobulins that reversibly precipitate at cold ▪ Hemolytic anemia, newborns, Prehepatic jaundice increased only
temperature. Cryoglobulins produce symptoms by precipitating in areas of low ▪ Bile duct obstruction, Post hepatic jaundice increased only
temperature, such as the skin or extremities, resulting in vascular occlusion. Inherited Bilirubin Derangements
Cryoglobulins are immunoglobulins that precipitate upon storage at refrigerated ▪ Deficiency of UDPGT CRIGLER NAJJAR
temperatures (4° C), and as a group, are associated with hematologic, ▪ Defective hepatic uptake of bilirubin, B1 GILBERT SYNDROME
autoimmune, and chronic infections such as hepatitis C. (Henry, 22nd ed) ▪ Defect in hepatic excretion of bilirubin, DUBIN JOHNSON
5) Methods for Proteins associated with dark liver pigmentation and SYNDROME
a. Measures amount of peptide bonds = BIURET REACTION abnormal gall bladder function; B2
Reagents = oc elle s salt l aline copper sul ate Na potassium iodide 13) Functions of Liver
Note: Rochelle's salt + Alk. Copper sulfate = alkaline copper tartrate a. Synthetic Function = Albumin, Proteins (except immunoglobulins),
b. Measures nitrogen content of proteins = Kjeldahl reaction coagulation proteins, cholinesterase, glycogen
Assumes that average nitrogen content of protein is 16% b. Metabolic Function = Conversion of glucose to acetyl-CoA; amino acid
Factor used to convert nitrogen content → protein concentration = .54 conversions, fatty acid metabolism, gluconeogenesis
c. Most Commonly used dye-binding technique = BROM CRESOL GREEN c. Detoxification = Bilirubin, Ammonia and drugs
d. Most specific and sensitive used dye-binding technique for albumin = d. Excretory Function = Bile acids excretion
BRCOM CRESOL PURPLE
e. Dye used for amino acids = NINHYDRIN
Enzymes
1) Enzyme class where LDH, glucose oxidase and G6PD OXIDOREDUCTASE
. Turbidimetric Methods for Protein = SULFOSALICYLIC ACID AND
belongs:
TRICHLOROACETIC ACID
Enzyme class where CK, AST, ALT and GGT belongs: TRANSFERASE
6) Total protein minus albumin N
Enzyme class where ACP, ALP, LPS, AMS , cholinesterase HYDROLASE
7) Total Bilirubin minus direct bilirubin N C N
belongs:
8) Icterus index involves diluting serum with saline until it visually matches the
Enzyme class of aldolase: LYASE
color of a 0.01% potassium dichromate solution. The number of times the
Factors that influence enzymes: Substrate concentration; Enzyme
serum must be diluted is called the icterus index. This method is subjective and
concentration; Temperature; pH, Inhibitors
substances in the serum other than bilirubin, such as carotene, xanthophyll and
. Mechanism of enzyme inhibition where the inhibitor binds the active site of
hemoglobin can contribute to icterus index.
the enzyme = C M N N

8
 b. Cofactors (coenzymes and activators) mylase Ca and Cl C M 9) Conditions associated with the greatest increase of enzymes:
c. Inactivation of CK can partially be reversed by addition of sulfhydryl LDH = pernicious anemia CK = uc enne s muscular dystrop y
compounds (i.e. N-acetylcysteine, mercaptoethanol, thioglycerol, and ALP = a et s disease AST/ALT = acute iral epatitis
dithiothreitrol) to the assay reagent 1st enzyme to be increased in AMI = CK-MB
3) Methods for measuring enzymatic reaction Increases after: ours Peaks after: ours
a. Fixed time method = reactants are combined, the reaction proceeds for a Normalizes within: days
designated time, after which the reaction is stopped by inactivating the CKPRONOUNCED ELEVATION MILD OR MODERATE ELEVATION
enzyme and then measurement of is made; the reaction is assumed linear (5 OR MORE TIMES NORMAL) (2-4 TIMES NORMAL)
over the reaction time (the larger the reaction, the more enzyme is present) Duchenne’s muscular Severe exercise, trauma, surgical
b. Continuous-monitoring (or kinetic assays) = involves multiple dystrophy procedure, intramuscular injections
Polymyositis Delirium tremens, alcoholic myopathy
measurements, usually of absorbance change, at specific time intervals or
Dermatomyositis severe ischemic injury
continuously via a continuous-recording spectrophotometer Myocardial infarction Pulmonary infarction
4) Enzyme Catalytic Reaction: E + S → ES → E + P Pulmonary edema (some patients)
5) Michaelis Menten Hypothesis: V = Vmax [S] / Km + [S] Hypothyroidism
Where: V = velocity of reaction S = Substrate concentration Acute agitated psychoses
Vmax = maximum velocity Km = Michaelis constant 12) Reference range of CK-MB: <6% of total CK
6) Enzyme kinetics where reaction rate is directly proportional FIRST ORDER 13) Distribution of LDH LD1 = heart, brain, RBCs
to the substrate concentration LD2 = heart, brain, RBCs
Enzyme kinetics where it is best to measure enzyme; ZERO ORDER LD3 = brain, kidney
reaction rate depends only on the enzyme concentration LD4 = liver, skeletal muscles, kidneys
7) Enzymes and disease associations LD5 = liver, skeletal muscles, ileum
▪ Acute myocardial infarction C M Cold labile LDH = and
▪ Acute pancreatitis mylase ipase LD Flip (LD1 > LD2) occurs in = C M C N C N
▪ Hepatic disease 16) Least organ specific enzyme = C N
▪ Hepatobiliary disease (liver and bile duct) 17) LDH
▪ Medico-legal evaluation of rape cases C PRONOUNCED ELEVATION MODERATE ELEVATION SLIGHT ELEVATION
▪ Skeletal muscle disorder ldolase C (5 OR MORE X N) (3-5 TIMES NORMAL) (UP TO 3 X N)
▪ Blood pressure regulation renin C Megaloblastic anemia Myocardial infarction Most liver diseases
Widespread carcinomatosis, Pulmonary infarction Nephrotic syndrome
8) Classifications of Enzymes
especially hepatic Hemolytic conditions Hypothyroidism
▪ Class 1 – oxidoreductase = LDH metastases Leukemias Cholangitis
▪ Class 2 – transferases = CK, GGT, AST, ALT Systemic shock and hypoxia Infectious mononucleosis
▪ Class 3 – hydrolases = ACP, ALP, Amylase, Lipase, Cholinesterase Hepatitis Delirium tremens
▪ Class 4 – lyase = aldolase Renal infarction Muscular dystrophy

9
Alpha-hydroxylbutryic dehydrogenase (HBD) has the same kinetic properties as
LD1 but does not act on the substrate lactate GGT, 5-nucleotidase, leucine aminopeptidase are enzymes that can differentiate
Cholinesterase catalyzes the hydrolysis of choline ester or acetylcholine into acid liver from bone elevation of ALP. They are increased in liver problems, but normal
and choline in bone-related diseases.
Increase of alkaline phosphatase concentration occurs after ingestion of HIGH GGT is a sensitive marker for alcoholism and hepatobiliary disease.
FAT MEAL. Amylase that is inhibited by wheat germ lectin = M
ALP isoenzymes Amylase = Methods
▪ Fastest toward the anode = ▪ Saccharogenic (sugar-generating)
▪ Most heat stable among the normal ALP = C N measures reducing sugars produced by hydrolysis of starch
▪ Most heat labile among the normal ALP = N ▪ Amyloclastic (starch-cutting or iodometric method)
▪ Inhibited by urea = N measures decrease in substrate (starch) concentration due to
Alkaline Phosphatase amylase activity; indicator used is iodine
PRONOUNCED ELEVATION MODERATE ELEVATION SLIGHT ELEVATION ▪ Coupled enzyme reaction or Continuous monitoring
(5 OR MORE x N) (3-5 x N) (UP TO 3 x N) coupling of several enzyme systems to monitor amylase activity
Bile duct obstruction Granulomatous or Viral hepatitis ▪ Chromogenic (color breakdown/chromolytic)
(intrahepatic or infiltrative diseases of Cirrhosis measures the increasing color from production of product coupled
extrahepatic) liver Healing fractures with a chromogenic dye
Biliary cirrhosis Infectious mononucleosis Pregnancy ( Substrate for Cherry-Crandall method for lipase = N
Osteitis deformans (Paget’s Metastatic tumors in placental ALP)
ipase is ery stable in serum, with negligible loss of activity at room
disease) bone Normal growth
Osteogenic sarcoma Metabolic bone diseases patterns in temperature for one week or for 3 weeks at 4°C.
Hyperparathyroidism (rickets, osteomalacia) children
Substrate for Bessey-Lowry-Brock method and Bowers-Comb method of alkaline Electrolytes and Trace Elements
phosphatase = p nitrop enyl p osp ate 1) Major extracellular cation SODIUM
Reference method for alkaline phosphatase analysis = C M 2) Major extracellular anion CHLORIDE
Most specific substrate for prostatic form of acid phosphatase = 3) Major intracellular cation POTASSIUM
M NM N 4) Major intracellular anion PHOSPHATE
AST 5) Primary counterion of sodium CHLORIDE
PRONOUNCED ELEVATION MODERATE ELEVATION SLIGHT ELEVATION 6) Electrolyte Panel/Routinely measured electrolytes Na, K, Cl, HCO3
(5 OR MORE x N) (3-5 x N) (UP TO 3 x N)
7) Primary Contributor to Osmolality Na
Acute hepatocellular Biliary tract obstruction Pericarditis
damage Cardiac arrhythmias Cirrhosis 8) Analyte that least likely to affect the anion gap POTASSIUM
MYOCARDIAL Congestive heart failure Pulmonary infarction 9) ANION GAP is the difference between unmeasured anions and unmeasured
INFARCTION Metastatic or primary Delirium tremens cations. It is used as a quality control tool for electrolyte testing.
Circulatory collapse (shock) tumor in liver Cerebrovascular 10) Roles of Potassium:
ACUTE PANCREATITIS Muscular dystrophy accident Transmission of nerve impulses; Myocardial rhythm and contractility
Infectious mononucleosis

10
11) Chloride is involved in maintaining osmolality, blood volume and electrical 16) Elevated Osmolality can be due to
neutrality. Dehydration; Hyperglycemia (DM); Hypernatremia; alcohol poisoning (i.e.
12) Methods for Chloride Determination ethanol, methanol, ethylene glycol, or isopropyl alcohol); uremia
▪ Amperometric-coulometric titration (principle of Cotlove (accumulation of toxins in the blood) and kidney damage; stroke, head
chloridometer) is a method using COULOMETRIC generation of silver ions trauma and shock; mannitol therapy
which combine with chloride to quantitate chloride concentration. Excess 17) Trace Elements
silver ions, which were not bound to chloride is used to indicate endpoint. ▪ Majority of Iron is found in M N
The amperometric and coulometric titration in chloride determinations ▪ First Step in Photometric Iron Methods C C N
suffer from halogen interference. Ions such as bromide, iodide, -CN, - ▪ Important Part of Glucose Tolerance Factor C M M
CNS, and -SH. ▪ Trace element found in thyroid hormones N
▪ Mercuric titration (principle of Schales and Schales method) is based on
the reaction of chloride ions to mercuric ions to form mercuric chloride. NPNs, KFT and Blood Gases
Excess mercuric ions are then made to react with DIPHENYLCARBAZONE in 1) NPNs according to concentration
order to form violet blue color. Urea > amino acids > uric acid > creatinine > creatine > ammonia
2) End products
▪ End product of protein metabolism
▪ End product of muscle metabolism C NN
▪ Colorimetric method (Method of Skeggs) uses mercuric thiocyanate and
▪ End product of purine metabolism C C
ferric nitrate to form ferric thiocyanate, which is a reddish colored complex
3) Azotemia = elevated urea level in the blood; can be classified as prerenal, renal
with a peak absorbance at 480nm.
and post renal azotemia
Uremia = also known as uremic syndrome; elevated urea level with renal
failure
▪ Ion-selective electrode using an ion exchange membrane selective for 4) Causes of Prerenal azotemia
chloride ions. Membrane used is a combination of silver wire coated with perfusion/flow of blood to the kidney; normal renal function; congestive heart
AgCl. failure, shock, hemorrhage, dehydration, decreased blood volume
5) BUN to Creatinine Ratio 10:1 to 20:1
13) Common methods used for calcium analysis use orthocresolphthalein 6) Factor used to convert BUN into urea concentration: BUN x 2.14
complexone (OCPC) or arsenzo III dye to form complex with calcium; as well as c r used c er ure c ce r i i BUN x .
Atomic absorption spectrophotometry 7) Purpose of nitroprusside in Berthelot rxn for BUN CATALYST
14) etany is a neuromotor irritability accompanied by muscular twitching and 8) Urea method that is inexpensive but lacks specificity C M C M
eventual convulsions; generally due to lo calcium le els, but may also 9) Simplest Type of Jaffe reaction COLORIMETRIC ENDPOINT
be caused by lo ma nesium. More specific and more preferred type of Jaffe COLORIMETRIC-KINETIC
15) Analyte with reciprocal relationship with calcium PHOSPHATE Reagents in Jaffe reaction NaOH + picric acid Fuller’s earth/Lloyd’s rgt

11
10) NPN that absorbs light at 293nm URIC ACID 23) Primary compensatory mechanism in:
11) Method for Uric acid that is affected by interferences ENZYMATIC METHOD Metabolic acidosis HYPERVENTILATION
caused by reducing substances (H2O2) Metabolic alkalosis HYPOVENTILATION
12) Condition associated with deficiency of hypoxanthine LESCH-NYHAN
guanine phosphoribosyl transferase SYNDROME Endocrinology
13) Refers to the deposits of uric acid crystals as sodium urates 1) Thyroid Gland
TOPHI
in great toe, ear lobe, and elbow and in other tissues etc. ▪ Produces thyroid hormones (T3 and T4) and calcitonin
14) Reye’s Syndrome and Hepatic coma (hepatic ▪ Alkaline phosphatase levels in hyperthyroidism = INCREASED
AMMONIA
encephalopathy) is associated with elevated levels of: Cholesterol and triglycerides level in hypothyroidism = INCREASED
15) Functions of Kidneys ▪ Test analyte that confirms conflicting thyroid test results = rT3
▪ Synthetic Function ▪ rT3 is formed form the deiodination of T4 in the = BLOOD
Erythropoietin (hormone for RBC production and maturation, stimulus: ▪ Disease characterized by = T3 THYROTOXICOSIS
tissue hypoxia), Renin (converts angiotensinogen to angiotensin I), decreased TSH, normal FT4, (PLUMMER’S DISEASE)
Prostaglandins increased FT3 and increased T3:
• Renin can increase blood pressure, increase plasma sodium, ▪ Hyposecretion of T4 in children = CONGENITAL
increase water retention results to: HYPOTHYROIDISM (CRETINISM)
Functions of Angiotensin II ▪ Increased TAG, increased = HYPOTHYROIDISM
❖ Regulates blood pressure cholesterol, cold intolerance,
❖ Vasoconstriction and Stimulates aldosterone production weight gain, inactivity:
▪ Metabolic Function = activation of vitamin D ▪ Exophthalmia, resting tachycardia, = HYPERTHYROIDISM
▪ Excretory Function = removal of ammonia (in the form of urea), uric acid, weight loss, increased appetite,
creatinine, toxic substances hyperactivity, heat intolerance:
16) Organs that control acid-base balance: LUNGS AND KIDNEYS ▪ T3U is directly proportional to T3/T4 levels
17) Effect of every 1°C of fever on: pCO2 3% increased ▪ T3U is inversely proportional to the amount of available binding sites in TBG
pO2 7% decreased ▪ T3U in hypothyroidism = decreased
18) Venous blood is usually 0.03 pH units lower than arterial blood. ▪ T3U in hyperthyroidism = increased
19) Ratio of Bicarbonate to carbonic acid = 20:1 ▪ Protein carriers for thyroid hormone: 70-75% = TBG
20) Normal pH is 7.35-7.45 20% = TBPA
21) Changes in results if a blood gas specimen is exposed pH = 10% = TBA
to room air pO2 = T3/T4 TSH
pCO2 = Primary hypothyroidism decreased increased
Secondary hypothyroidism decreased decreased
22) Anticoagulant of choice for blood gas analysis = heparin in syringe with
Primary hyperthyroidism increased decreased
rubber stopper (do not use vacutainer tube!)
Secondary hyperthyroidism increased increased

12
 4) Three screening tests are used in the evaluation of a patient suspected of having
2) Sex Hormones and Pregnancy-related hormones
Cushing’s syndrome: the 24-hour urinary free cortisol, the overnight
▪ Most potent estrogen es r di
dexamethasone suppression test (DST), and the plasma or salivary midnight
▪ Predominant estrogen in menopausal women es r e
cortisol (MSC) level. (p391 Henry 23rd ed)
▪ Predominant estrogen in pregnant women es ri
5) Pancreas
▪ Estrogen type involved in menstrual cycle es r di
▪ Hormone that promotes glycolysis, glycogenesis and lipogenesis N N
▪ Primary source of estradiol ries
▪ Fragment removed from proinsulin in order to activate it C
▪ Primary source of estriol ce
6) Pituitary Gland
▪ Hormone increased during the 1st trimester of pregnancy;
▪ Anterior Pituitary Gland (Adenohypophysis) produces:
serves as marker of Pregnancy; marker of testicular
ACTH, TSH, FSH, LH, Prolactin and Growth hormone
carcinoma, produced by the placenta
▪ Hypersecretion of growth hormone in adulthood can cause C M
▪ Hormones increased in the 2nd and 3rd trimester of pregnancy r es er e
▪ Hypersecretion of growth hormone in childhood can cause N M
▪ Most potent male sex hormone es s er e
▪ Glucose suppression test is used for suspected cases of acromegaly.
▪ Condition associated with:
▪ Storage site for ADH and oxytocin = POSTERIOR PITUITARY GLAND
High FSH & LH, low testosterone = Primary deficiency of male sex hormones
▪ Target sites of ADH/vasopressin action = DCT and CD
Low FSH & LH, low testosterone = Secondary deficiency of male sex hormones
▪ Stimulus for ADH/vasopressin release = HYPEROSMOLAR PLASMA
High FSH & LH, low E2 = Primary deficiency of female sex hormones
7) Substances that regulate calcium: Parathyroid hormone, Vitamin D, Calcitonin
Low FSH & LH, low E2 = Secondary deficiency of female sex hormones
8) Primary metabolite of serotonin = 5-HIAA ydr yi d e ce ic cid
Increased hCG: Down Syndrome, Choriocarcinoma, Molar pregnancy
Decreased hCG: Ectopic pregnancy TDM and Toxicology
▪ Chromosomal Abnormalities (Trisomies) Function of Therapeutic drugs
Patau syndrome = Trisomy 13 ▪ Used for treatment of manic-depression (bipolar disease) = M
Edward syndrome = Trisomy 18 ▪ Bronchodilator/ anti-asthma drug = eop yline
Down syndrome = Trisomy 21 ▪ Barbiturates and benzodiazepines are the most common type of sedative
Adrenal glands hypnotics abused. Secobarbital, pentobarbital and phenobarbital are the more
▪ Substance that promotes vasoconstriction, stimulates = commonly abused barbiturates.
aldosterone production and regulates blood pressure ▪ Some of the anticonvulsants (antiepileptic) drugs include: valproic acid,
▪ Condition associated with hypercortisolism, moon- = H phenobarbital, carbamazepine and phenytoin.
shaped face and buffalo hump ▪ Anticonvulsant that can be used for grand mal seizure (tonic-clonic) =
▪ Major mineralocorticoid = N
Major glucocorticoid = ▪ Anticonvulsant that can be used for petit mal (absence) seizures = C C
▪ i y de ic cid is e ie = H ▪ Associated to the red man syndrome; used for MRSA = VANCOMYCIN
▪ H i ic cid is metabolite = ▪ Antibiotic associated with aplastic anemia = CHLORAMPHENICOL
▪ Function of cyclosporine = IMMUNOSUPPRESSANT

13
 ▪ Function of Digoxin = CARDIOACTIVE or ANTI-ARRHYTHMIC DRUG 14) Specimen for Lead Toxicity = H
▪ Codeine, anti its analog dextromethorphan is used as a mild analgesic and an 15) Most commonly abused substance = H
antitussive agent (cough suppressant) 16) Blood Alcohol Levels
2) Serum drug concentration is affected by absorption, distribution and metabolism BLOOD ALCOHOL
SIGNS AND SYMPTOMS
3) Specimen collected before the administration of next dose = (% w/v)
4) Acetaminophen is toxic to which organ? = No obvious impairment, some changes observable on
0.01 – 0.05
5) Trinder’s reagent, which uses ferric nitrate is used in = C performance testing
assessment of toxicity with: Mild euphoria, decreased inhibitions, some impairment of
0.03 – 0.12
6) The techniques for identifying and measuring the concentration of small amounts motor skills
of drugs in such a complex protein matrix as serum are varied and sometimes Decreased inhibitions, loss of critical judgment, memory
0.09 – 0.25
intricate. No single instrument or technique can be used for all the different types impairment, diminished reaction time
of drug to be measured. Methods include: Mental confusion, dizziness, strongly impaired motor
0.18 – 0.30
a. Enzyme immune assay systems skills (staggering, slurred speech)
-Enzyme multiplied immunoassay technique (EMIT) UNABLE TO STAND OR WALK, VOMITING, IMPAIRED
0.27-0.40
-Enzyme-linked immunosorbent assay (ELISA) CONSCIOUSNESS
-Enzyme immunochromatography 0.35-0.50 COMA AND POSSIBLE DEATH
b. Fluorescence immunoassay PRESUMPTIVE EVIDENCE FOR DRIVING UNDER
≥ 0.10
-FPIA (i.e. TDX) INFLUENCE OF ALCOHOL (LEGALLY INTOXICATED)
c. Radioimmunoassays (RIA)
d. Chromatography
-High performance liquid chromatography (HPLC)
-Gas-Liquid chromatography (GLC)
-Thin-layer chromatography (TLC)
e. Spectrophotometry
-Atomic Absorption Spectrophotometry (AAS)
7) Metabolite of Cocaine = BENZOYLECGONINE
8) Metabolite of heroin = MORPHINE
9) Major active component of Marijuana = Δ-9-TETRAHYDROCANNABINOL
10) Confirmatory Test for Drug Analysis = GC-MS
11) Odor of garlic; Keratinophilic, hair, nails and skin can be used as specimen to
determine toxicity; found in insecticides and pesticides = NC
12) Odor of Bitter almonds =C N
13) Has high affinity to hemoglobin, (200-245x); primary sources are car exhausts
and cigarette smoke; cherry red color of blood =

14
 <3.> Specimen Collection and Considerations
MICROBIOLOGY + PARASITOLOGY a. Blood
▪ collected 30-45minutes before the fever spikes
<1.> Stages of Bacterial Growth ▪ 2-3 sets of blood cultures per 24 hours
. Adjustment phase = LAG PHASE b. Sputum
. stage where the bacterial cells divide at a constant rate; stage where the ▪ Early morning specimens collected on two -three consecutive days
organisms are most active metabolically; stage where the organisms are most ▪ First thing to be done before collection of sputum samples: gargle with
susceptible to antibiotics = / N N water
c. Stage where the number of dying organisms is equal to the number of viable ▪ Specimens with too many squamous epithelial cells (>10/hpf) and/or too
organisms = N few PMN cells (<25/hpf) are not suitable for culture
d. Stage where the number of dying organisms is greater than the number of viable ▪ Inoculating loops and needles used for sputum culture should be dipped in
organisms = / C N 70% alcohol or cresol with sand before flaming
<2.> Nutritional Requirements of Bacteria ▪ N-acetyl-L-cysteine (NALC) = Digestion
a. Obligate Aerobes ▪ 2-4% NaOH = Decontamination
Oxygen = Carbon dioxide = 0.03% c. Specimen for detection of carriers of Staphylococcus aureus = N
. Capnophiles d. Preferred specimen for Bordetella pertussis; detects carriers of Neisseria
Oxygen = 15% Carbon dioxide = meningitidis = N N
c. Microaerophiles e. Specimen for detection of Streptococcus pyogenes (strep throat), Candida albicans
Oxygen = 5% Carbon dioxide = 10% Nitrogen = 85% (oral thrush), Haemophilus influenzae (epiglottitis) =
d. Anaerobic Systems of Incubation . Urine
Anaerobic jar Evacuation-replacement system ▪ Significant bacteriuria = >105 CFU/mL (or 100,000)
Glove-box method Disposable anaerobic bags ▪ Midstream clean catch; first morning urine is preferred since it is more
brewer jar Torbal jar, Gas Pak jar concentrated
NOTE: Candle ar is N an anaerobic ar. >___< g. Stool
e. Characteristics of Anaerobic Incubation ▪ 3 specimens, one per day for 3 days should be collected
▪ Palladium pellets are used as catalysts h. Wounds and Abscesses
▪ Indicator of effectiveness of anaerobic system: M N ▪ Needle and syringe aspirate enhances the recovery of anaerobes
f. Carbon source is from carbon dioxide and other inorganic substances = <4.> Stains
AUTOTROPHS/ LITOTROPHS a. Schaeffer-Fulton Stain = endospores
g. Carbon source is from organic compounds like carbohydrates, lipids and b. Leifson Stain = flagella
proteins = HETEROTROPHS/ ORGANOTROPHS c. Methylene blue = Volutin or Babes-Ernst or Metachromatic Granules
h. Organism that grows both at 35°C and 42°C = Pseudomonas aeruginosa, d. Wayson = Yersinia
Campylobacter e. India Ink = negative stain; capsules of Cryptococcus

15
 f. Gram Stain New York City agar = CAP + vancomycin-colistin-amphotericin B-trimethoprim
▪ Primary Stain = crystal violet lactate
▪ Mordant = Gram’s iodine e. Bismuth sulfite agar = almonella
▪ Decolorizer = acetone alcohol . Regan Lowe (charcoal cephalexin blood) agar = ordetella pertussis
▪ Secondary Stain = safranin O . Cystine-tellurite medium, Potassium-tellurite medium, modified Tinsdale
▪ Most critical step in Gram staining technique = decolorization medium = Corynebacterium dip t eriae
▪ Gram positive = purple-blue Gram negative = pink-red . Blood Cystine Glucose agar = Francisella tularensis
▪ Gram ghost = Mycobacterium i. Buffered Charcoal Yeast extract = Legionella pneumophila
▪ Pleomorphism = variability in the size and shape of cells . Skirrow’s medium = Campylobacter sp
g. Acid-Fast Stain . Thiosulfate citrate bile salt sucrose medium (TCBS) = ibrio sp.
▪ Ziehl-Neelsen Method = hot method; Kinyoun Method = cold method . MacConkey sorbitol agar = Escherichia coli O157:H7
▪ Acid fast organisms in tissues are best stained using = Kinyoun . Human Blood Tween (HBT) Bilayer medium = ardnerella a inalis
▪ Primary Stain = carbol fuchsin . Middlebrook 7H10 and 7H11 = Mycobacterium
▪ Decolorizer = acid alcohol (95% ethanol + hydrochloric acid) . Todd-Hewitt broth is recommended for CULTURE OF BETA-HEMOLYTIC
▪ Secondary Stain = methylene blue or malachite green STREPTOCOCCI FOR FLUORESCENCE MICROSCOPY
Hematoxylin as counterstain = Fite-Faraco method <6.> Colonial Morphology of Certain organisms
▪ Acid Fast organisms = RED; Non-acid fast organisms = BLUE . MacConkey
▪ Diameter of sputum in the slide during AFB smear preparation = 2-3cm ▪ Lactose fermenters = pin red Non-lactose fermenters = colorless
<5.> Culture Media . Eosin-Methylene blue medium
. Blood Agar Plate ▪ Greenish metallic sheen = sc eric ia coli
▪ 5% defibrinated blood; Choice of blood: Sheep, Horse, Rabbit c. Xylose-Lysine Desoxycholate (XLD) agar
▪ Hemolytic patterns ▪ Yellow = carbohydrate fermenters
o Greenish zones of hemolysis, incomplete hemolysis = Alpha ▪ Red with black centers = Salmonella
o Clear zones of hemolysis, complete hemolysis = Beta d. Cefsulodin Irgasan Novobiocin Medium
o Small zone of alpha surrounded by a zone of beta hemolysis after ▪ Bull’s eye colonies = Yersinia enterocolitica
refrigeration = Alpha prime <7.> Indicators found in certain culture and biochemical media
. Mannitol salt agar = y c ccus s . e. Phenol red = XLD; TSI; urease test, Mannitol Salt Agar
c. Chocolate agar = Neisseria, Haemophilus . Methyl red = MR test
d. Thayer Martin = CAP + vancomycin-colistin-nystatin . Neutral red = MacConkey agar
Modified Thayer Martin = CAP + vancomycin-colistin-nystatin-trimethoprim . Bromthymol blue = Malonate utilization test; Simmon’s Citrate Agar;
lactate Carbohydrate Fermentation Test (OF test)
Martin-Lewis Medium = CAP + vancomycin-colistin-anisomycin-trimethoprim i. Bromcresol purple = Lysine Iron Agar
lactate

16
<8.> Biochemical Tests . Oxidase test
. Motility ▪ Reagent: 1% tetramethyl para-phenylene diamine dihydrochloride
▪ Motile at r 25°C but not at 37°C = Yersinia enterocolitica ▪ Positive Result: Maroon-violet or dark-purple color after 10-30secs
▪ Klebsiella, Shigella = NON-MOTILE ▪ Procedure:
. IMVIC Reactions of some organisms o Use platinum loop or wooden applicator to remove colony; presence of
▪ Escherichia coli = (+ + 0 0) iron in the nichrome loop can give false positive reaction
▪ Klebsiella pneumoniae and Enterobacter aerogenes = (0 0 + +) o Rub the colony or a paper strip and add reagent or rub the colony on a
c. Substrate used for indole test: TRYPTOPHAN piece of paper containing the reagent or put a drop of reagent in colony
d. Test to differentiate Klebsiella from Enterobacter = ORNITHINE
DECARBOXYLASE and MOTILITY . Phenylalanine Deaminase test
e. Test to differentiate Klebsiella oxytoca from Klebsiella pneumoniae = INDOLE ▪ Reagent: 10% ferric chloride and phenylalanine agar
Test to differentiate Proteus vulgaris from Proteus mirabilis = INDOLE TEST ▪ Positive: Proteus, Providencia, Morganella
f. Biochemical reaction with reciprocal relationship = METHYL RED and VOGES- . Cetrimide Test
PROSKAUER ▪ Checks ability of organism to grown in the presence of toxic cetrimide
g. Methyl Red Test ▪ Positive: seudomonas aeru inosa
▪ Detects for mixed acid production; positive result: RED . Acetamide Utilization Test
h. Voges-Proskauer ▪ Checks the ability of an organism to use acetamide as sole source of carbon
▪ Detects acetoin or acetyl methyl carbinol ▪ Bacteria can deaminate acetamide to release ammonia, causing a shift in
▪ Reagent: KOH and alpha-naphthol alkaline pH
▪ Positive result: red ▪ Change in color from green to blue
i. Triple Sugar Iron Agar ▪ Do not inoculate organism from a broth culture, since it can cause
▪ Ratio of lactose, sucrose and glucose = 1: 1: 0.1 heavy growth
▪ H2S indicator = ferrous ammonium sulfate ▪ Incubate at de C or days Positive: seudomonas aeru inosa
j. ONPG Test . Catalase Test
▪ Detects for the presence of beta-galactosidase ▪ Reagent: hydrogen peroxide (usually 3%)
▪ Differentiates late lactose fermenters from non-lactose fermenters ▪ Positive result: bubbles or effervescence, gas production
k. Lysine-Ornithine-Arginine (LOA) activity of Enterobacter species . Superoxol Test
Enterobacter aerogenes ++0 ▪ Presumptive test for Neisseria onorr oeae
Enterobacter cloacae 0++ ▪ Reagent: 30% hydrogen peroxide; positive result = bubblin
Enterobacter gergoviae ++0 r. Coagulase Test
Enterobacter sakazakii 0++ ▪ Reagent: rabbit plasma
Enterobacter taylorae 0++
▪ Slide coagulase = detects bound coagulase (clumping factor)
▪ Tube coagulase = detects free coagulase
▪ Positive: Staphylococcus aureus

17
s. DNAse test (DNA hydrolysis test) <9.> Serological Tests
▪ Positive Result: Methyl green = green to colorless a. For Diagnostic testing
Toluidine blue = blue to rose pink ▪ Schultz-Charlton = Streptococcus pyogenes
▪ Positive: Staphylococcus aureus, Moraxella catarrhalis, Serratia ▪ Widal test = Salmonella
t. CAMP Test ▪ Weil-Felix test = Rickettsia
▪ Positive result: arrow head zone of beta hemolysis Reagent antigens: OX-K from Proteus mirabilis
▪ Negative result: no enhancement of hemolysis OX-2 and OX-19 from Proteus vulgaris
u. Neufeld Quellung Test (Capsular Swelling Test) ▪ Casoni skin test = Echinococcus granulosus
▪ Quellung = swelling ▪ Frei test = lymphogranuloma venereum
▪ Positive: organisms with capsule (e.g. Streptococcus pneumoniae) b. For Susceptibility testing
v. Bile Solubility test ▪ Mantoux, Mendel’s, Tuberculin, Vollmer’s, von Pirquet = tuberculosis
▪ Positive Result: colony lysis; Positive: Streptococcus pneumoniae ▪ Schick’s test = diphtheria
w. Acetate Test ▪ Dick’s test = scarlet fever
▪ Checks the ability of an organism to use acetate as sole source of carbon c. For Bacteria classification
▪ Bacteria can breakdown acetate, causing a shift in alkaline pH ▪ Lancefield Classification = Streptococcus; based on the specific cell wall
▪ Change in color from green to blue polysaccharide (cell wall antigen)
▪ Do not inoculate organism from a broth culture, since it can cause Capsules are used for serotyping
▪ Kauffman-White scheme = Salmonella
heavy growth
<10.> Xenodiagnosis: Ocular Test of Anton = LISTERIA MONOCYTOGENES
▪ Incubate at 35°C for up to 7 days
<11.> Microbial Control
▪ Positive: Escherichia coli
. Iodophor = iodine deter ent (e.g. N )
x. MUG test
. Iodine tincture = 2% iodine + 50% alcohol
▪ Detects the presence of beta-D-glucuronidase
c. For sterilizing glassware = dry heat oven (160-180°C for 1.5-2 hours)
▪ Positive: Escherichia coli
d. Autoclave
▪ Positive result: electric blue fluorescence
▪ moist heat sterilization, 121°C for 15 minutes at 15psi
▪ Negative result: lack of fluorescence
▪ Indicator: Bacillus stearothermophilus
y. Carbohydrate Utilization Test for Neisseria
e. Media that cannot be autoclaved can be: FILTERED
▪ N. gonorrhoeae–glucose only
. Fractional Sterilization
▪ N. meningitidis–glucose and maltose
▪ Tyndallization – 100°C for 30 minutes, 3 consecutive days
▪ N. lactamica–glucose, maltose & lactose
▪ Inspissation – 75-80°C for 2 hours, 3 consecutive days
▪ N. sicca-glucose, maltose, fructose & sucrose
. Pasteurization
z. Chromogenic beta lactamase test uses Cefinase disks. Positive result is change
▪ Batch = 63°C for 30 minutes
in color from yellow to red. (change in color/color formation)
▪ Flash = 72°C for 15 sec
aa. In vitro toxigenicity test for Corynebacterium diphtheriae = Elek test

18
 h. Sodium Hypochlorite (NaOCl, Household bleach; 1:10 dilution) e. Novobiocin
▪ Inactivates: HBV within 10 minutes Sensitive: Staphylococcus epidermidis
HIV within 2 minutes Resistant: Staphylococcus saprophyticus
i. Quaternary Ammonium Compounds (e.g. Benzalkonium chloride) <15.> Antibiotic interactions:
▪ Inactivated by organic substances . Autonomous/Indifferent ~ results obtained with two drugs is equal to result
j. All laboratories must adhere to guidelines of BIOSAFETY LEVEL 2 with most effective drug by itself
k. Most hospital microbiology laboratories routinely use . Antagonistic ~ result with two drugs is significantly less than the autonomous
CLASS II-A BIOLOGICAL SAFETY CABINETS result
l. Benchmark is a reference point. Benchmarking is seeking and industry or c. Additive ~ result with two drugs is equal to the combine action of each of the
profession’s best practices to imitate and improve. It involves evaluation or drugs used separately
checking by comparison with a standard. d. N C ~ result with two drugs is significantly greater than
<12.> Antibiotics and their action
additive response
a. Penicillin and Vancomycin = INHIBITS CELL WALL SYNTHESIS
<16.> Antimicrobial Susceptibility Testing (Kirby-Bauer)
b. Quinolones, Rifampin, Sulfonamides = INHIBITS DNA SYNTHESIS
. Medium used: Mueller Hinton Agar (MHA)
c. Aminoglycosides (Gentamicin), Clindamycin Chloramphenicol = INHIBIT
. pH of MHA = . . Thickness or depth of MHA = mm
PROTEIN SYNTHESIS
c. Bacteria = equivalent to 0.5 McFarland Standard which contains
<13.> Antibiotic Treatment
approximately . C /m bacteria
a. For Tuberculosis, causes red-orange discoloration of urine and body fluids =
<17.> Bacteria and their common names
Rifampin
. Friedlander’s bacilli = e sie eu i e
b. For leprosy = Sulfone dapsone
. Pfeiffer’s bacilli = Haemophilus influenzae
c. For MRSA = vancomycin
c. Koch-Week’s bacilli = Haemophilus aegyptius
d. ESBL stands for Extended Spectrum Beta Lactamase
d. Koch’s bacilli = yc c eriu u ercu sis
<14.> Bacteria and Antibiotics
e. Tap water bacilli = Mycobacterium gordonae
a. Furazolidone
. Canned good bacilli = Clostridium botulinum
Sensitive: Staphylococcus Resistant: Micrococcus
. Fried rice bacilli = Bacillus cereus
b. Lysostaphin: <18.> Bacteria and their pathogenic determinants
ensiti e tap ylococcus Resistant: Micrococcus
a. Gram positive organisms usually produce = exotoxins
c. Bacitracin (Taxo A, . units) Gram negative organisms usually produce = endotoxins
Sensitive: Micrococcus and Streptococcus pyogenes
b. Most potent toxin = Botulinum toxin (Clostridium botulinum)
Resistant: Staphylococcus
c. Staphylococcus aureus
d. Optochin: (Taxo P; ethyl hydrocuprein hydrochloride)
▪ Protein A
Sensitive: Streptococcus pneumoniae
▪ Panton-Valentin Factor = leukocidin
Resistant: Viridans streptococci
▪ Duran-Reynal Factor = hyaluronidase; spreading factor
▪ Exfoliative toxin = responsible for scalded skin syndrome

19
 d. Streptococcus pyogenes (Group A, beta hemolytic Streptococci)
▪ Streptolysins (hemolysins)
Streptolysin O Streptolysin S
Oxygen-labile Oxygen-stable
Subsurface hemolysin Surface hemolysin
immunogenic Non-immunogenic
e. ETEC (Enterotoxigenic E. coli) → Traveler’s diarrhea
▪ Choleragen-like toxin
f. EHEC (Enterohemorrhagic E. coli)
▪ Shiga-like toxin
<19.> Bacteria: disease associations and characteristics
▪ Golden yellow colonies in BAP, grape-like clusters, pinhead colonies =
Staphylococcus aureus
▪ Toxic shock syndrome, scalded skin syndrome = Staphylococcus aureus
▪ UTI in young females = tap ylococcus saprop yticus
▪ Bacterial endocarditis after prosthetic heart valve insertion = tap . epidermidis
▪ Tests to differentiate S. aureus from S. epidermidis = coagulase and DNAse test
▪ Gram positive cocci in chains, pinpoint colonies = Streptococci
▪ Primary normal flora of oral cavity; associated with subacute bacterial
endocarditis = VIRIDANS STREPTOCOCCI
▪ Strep throat (acute pharyngitis), scarlet fever, erysipelas, impetigo, cellulitis,
PSAGN and acute rheumatic fever = Streptococcus pyogenes
▪ Neonatal meningitis, beta hemolytic Group B, CAMP positive and hippurate
hydrolysis positive = Streptococcus agalactiae
▪ Lobar pneumonia, rusty red or anchovy red sputum = Streptococcus
pneumoniae
▪ Bile esculin test positive; 6.5% salt turbidity test positive = Group D
Enterococcus
▪ Bile esculin test positive; 6.5% salt turbidity test negative = Group D non-
Enterococcus
▪ Nutritionally-variant Streptococci require Vitamin B6 or cysteine
▪ Ophthalmia neonatorium: Neisseria gonorrhoeae
▪ Specimen for N. gonorrhoeae can be taken from: eyes, rectum, oral cavity
▪ Most common cause of urinary tract infection = Escherichia coli
▪ Biochemically inert member of Enterobacteriaceae family = Shigella
▪ Swarming colonies, gun powder odor = Proteus
▪ Agent of plague, associated with bubo, transmitted by the flea Xenopsylla
cheopis, Stalactite patterns in broth culture = ersinia pestis
▪ Flat, spreading, serrated edges (feathery edges) in blood agar plate with grape-
like or fruity odor, produces pyocyanin and pyoverdin; polar flagellum =
Pseudomonas aeruginosa
▪ Causative agent of swimmer’s ear = seudomonas aeru inosa
▪ Causative agent of Melioidosis = Burkholderia (Pseudomonas) pseudomallei
▪ Glander’s disease = Burkholderia (Pseudomonas) mallei
▪ Non-halophilic Vibrio sp. = Vibrio mimcus and Vibrio cholerae
▪ Associated with rice watery stool = Vibrio cholerae
▪ Associated with septicemia and wound infections in patients handling seafood
or working in marine environment with pre-existing liver disease = Vibrio
vulnificus
▪ Blood-loving organisms on CAP = Haemophilus
▪ Most common cause of epiglottitis = Haemophilus influenzae
▪ Associated with chancroid or soft chancres, schools of fish in Gram stain = . ducreyi
▪ Hemophilus species (Delost)
Requires X and V = H. influenzae, H. haemolyticus, H. aegyptius
Requires X only = H. ducreyi
Requires V only = H. parainfluenzae, H. parahaemolyticus, H.
paraaprophilus
Does not require X and V = H. aprophilus
▪ Darting motility; Seagull wings appearance = Campylobacter sp.
▪ Associated with peptic ulcers, strongly urease positive and microaerophilic =
Helicobacter pylori
▪ Associated with whooping cough; mercury droplets in Regan Lowe = Bordetella
pertussis
▪ Stages of Pertussis =- catarrhal, paroxysmal, convalescent phase
▪ Differentiation of Bordetella species (Delost)
B. pertussis B. parapertussis B. bronchiseptica
Urease 0 + (slow) + (rapid)
Oxidase + 0 +
20
▪ Causative agent of Pontiac Fever or Legionnaire’s disease, isolated from air
conditioning towers and heating systems = Legionella pneumophila
▪ Can cause bacterial vaginosis, associated with the presence of clue cells in urine
and has a positive Whiff (KOH) Test = Gardnerella vaginalis
▪ Capnophilic organism with gliding motility = Capnocytophaga
▪ Causes granuloma inguinale = Klebsiella Calymmatobacterium)
▪ uses swimming pool granuloma = Mycobacterium marinum
▪ Bleach-like odor, capable of pitting the agar = Eikenella corrodens
▪ Associated with animal scratches and bite; with typical musty odor or odor of
mushroom = Pasteurella multocida
▪ Produces violet pigment known as violacein = Chromobacterium violaceum
▪ Capnophilic organisms associated with endocarditis: HACEK
Haemophilus aprophilus Eikenella corrodens
Actinobacillus mycetemcomitans Kingella denitrificans
Cardiobacterium hominis
▪ Gram negative coccobacilli commonly confused with Neisseria; oxidase negative
and obligate aerobe; grows in EMB and Mac = Acinetobacter sp.
▪ Pleomorphic, with Chinese character appearance, and metachromatic granules =
Corynebacterium sp.
▪ Corynebacterium diphtheriae produces GRAY BLACK colonies in Potassium
tellurite medium
▪ Listeria monocytogenes is bile esculin hydrolysis positive and catalase
positive.
▪ Tumbling end-over-end motility when incubated in Nu rie r at RT for
1-2 hours; umbrella-shaped pattern after overnight incubation at r
e er ure in a semi-solid medium; can be cultured on Cold enrichment
medium = Listeria monocytogenes
▪ Test tube brush growth in semi-solid medium, gram positive bacilli and H2S
positive; causes the Butcher’s disease, erysipeloid, seal finger or whale finger =
Erysiphelothrix rhusiopathiae
▪ Partial acid fast, Gram positive branching organism = Nocardia
▪ MPT64 is used for detection of = Mycobacterium tuberculosis
▪ Niacin positive, Nitrate positive = Mycobacterium tuberculosis
▪ Niacin negative, nitrate negative = Mycobacterium bovis
▪ Found in BCG (Bacillus of Calmette and Guerin) = Mycobacterium bovis
▪ Mycobacterium that is Tween 80 hydrolysis test positive = . s sii d .
rd e
▪ Associated with the presence of Lepra cells (macrophages containing acid-fast
bacilli); Causes Hansen’s disease = Mycobacterium leprae
u y s ssi ic i yc c eriu er
u ercu sis divides the Mycobacteria into photochromogens,
scotochromogens, non-chromogens and rapid growers.
▪ Bamboo pole/Fishing rod appearance on Gram stain, poly-D-glutamic acid on cell
wall, Medusa head colonies on sheep blood agar, non-hemolytic, non-motile;
causes Woolsorter’s disease or respiratory anthrax but also has cutaneous and
gastrointestinal (most dangerous) forms, agent of bioterrorism = Bacillus
anthracis
▪ Also known as Fried rice bacilli; hemolytic and motile = Bacillus cereus
▪ Most common laboratory contaminant = Bacillus subtilis
▪ Aerobic, Gram positive spore-forming rod = Bacillus sp.
▪ Anaerobic, Gram positive spore-forming rod = Clostridium sp.
▪ Gram positive, anaerobic cocci = Peptococcus and Peptostreptococcus
▪ Anaerobic infections are characterized by foul-smelling discharge
▪ Clindamycin-associated pseudomembranous colitis; cultured using CCFA
(cycloserine-cefoxitin-fructose agar) = s ridiu di ici e
▪ Lecithinase positive, double zones of hemolysis, reverse CAMP positive, Nagler
plate, causes gas gangrene or myonecrosis = Clostridium perfringens
▪ Tennis racket or drumstick appearance with round, terminal spores; causes Lock
jaw (Risus sardonicus) = Clostridium tetani
▪ The disease botulism results from ingestion of preformed toxin in nonacidic
vegetable or mushroom foodstuffs. Absorption of the toxin leads to nearly
complete paralysis of respiratory and other essential muscle groups. Other forms
of botulism can occur when the organism elaborates the toxin after it has
colonized the gastrointestinal tract of infants (i.e infant botulism). Wound
botulism is more rare than the other forms and occurs when Clostridium
botulinum produces the toxin from an infected wound site. (p460, Bailey &
Scott’s Diagnostic Microbiology 12th edition)
▪ Lumpy jaw, Molar Tooth colonies, Sulfur granules = Actinomyces
21
 ▪ Predominant normal microbiota of the gut = Bacteroides
▪ ▪ Certified milk
▪ Causative agent of Whipple’s disease = Tropheryma whipplei
o Contains 10,000 or less bacteria per mL
▪ Causes syphilis, characterized by hard chancres = Treponema pallidum
▪ Causes Lyme disease and erythema chronicum migrans, transmitted by ticks b. Bacteria found in milk
(Ixodes sp) = Borrelia burgdorferi ▪ Pseudomonas syncyanea = Blue milk
▪ Causes relapsing fever, transmitted by louse (Pediculus humanis) = Borrelia ▪ Serratia marcescens = Red milk
recurrentis ▪ Pseudomonas aeruginosa = Blue green milk
▪ Borrelia = loosely coiled; Leptospira = tightly coiled ▪ Clostridium butyricum and Clostridium perfringens = stormy
▪ Causes Weil’s disease = Leptospira interrogans icterohaemorrhagiae fermentation of milk
▪ Confirmatory test for Leptospira: culture ▪ Flavobacterium synxanthum = Yellow milk
▪ Chlamydia, Rickettsia and Mycoplasma contain both DNA and RNA. ▪ Alcaligenes viscosus = Slimy or ropy milk
▪ Rickettsia
c. Phosphatase Test – used to check effectiveness of pasteurization
Rickettsia rickettsii Causes Rocky mountain spotted fever ticks
▪ Phosphatase activity is always minimal in milk that has been completely
Rickettsia akari Causes rickettsial pox mites
Rickettsia Causes epidemic typhus, sporadic typhus, lice pasteurized
prowazekii Brill-Zinsser disease, flying squirrel typhus ▪ Methylene blue is added to milk
Rickettsia typhi Causes murine typhus, endemic typhus, flea- fleas ▪ Dye is decolorized (reduced) by oxygen consuming bacteria in sample
borne typhus ▪ Speed of decolorization is directly proportional to number of bacteria
Orientia Causes scrub typhus chigger ▪ Grading of Milk
tsutsugamushi CLASS MILK RATING DECOLORIZATION TIME
Bartonella Causes Oroya fever, Verruga peruana Lutzomiya I Excellent Over 8 hours
bacilliformis sandfly bite
II Good 6-8 hours
Bartonella henselae Causes cat-scratch disease, bacillary kitten or
III Fair 2-6 hours
angiomatosis and peliosis, endocarditis scratch bites
IV Poor <2 hours
Bartonella quintana Causes Trench fever or endocarditis lice
<21.> Water Bacteriology
(Pediculus)
▪ Causes Q fever, usually transmitted through aerosols or by ticks = ________________ a. Presumptive Test
▪ Organism that has two forms: an infectious elementary body and a replicating ▪ Media used: lactose broth or lauryl tryptose broth
reticulate body = Chlamydia sp. ▪ Positive result: gas production
▪ Causes lymphogranuloma venereum = Chlamydia trachomatis Addt’l info: Durham tube – can be used to detect gas production
▪ Smallest free-living organism that lacks cell wall = Mycoplasma b. Confirmed Test
<20.> Milk Bacteriology ▪ Media used: Endo agar
a. Milk Grades:
Brilliant green lactose bile broth (BGLBB)
▪ Grade A milk
o Obtained only from tuberculin negative cows Levine’s Eosin-Methylene blue agar plates
o Bacterial count below 75,000 bacteria/mL when raw ▪ (+) in BGLBB – gas formation within 48 hours
o Bacterial count should not exceed 15,000 bacterial/mL when ▪ (+) in Endo agar or Levine’s EMB agar – typical or atypical colonies w/in 24
pasteurized hrs

22
 c. Completed Test
▪ Media used: lactose broth fermentation tube and agar slant
▪ (+) Formation of acid and gas in lactose broth
▪ (+) demonstration of gram negative non-sporeforming bacilli in the agar is
considered positive
<22.> Parasitology
. Oviparous parasite = parasites that lays or produces eggs
Larviparous or viviparous parasites = parasites that produces larva (e.g.
Trichinella, Filarial worms)
. Mode of Transmission
▪ Hookworms, Strongyloides, Schistosomes = N N N specimen of choice is skin snips
▪ Ascaris, Trichuris, Entamoeba, Giardia = INGESTION
▪ Toxoplasma = CONGENITAL
c. Nematodes/Roundworms
▪ Unholy Three = Hookworms, Ascaris lumbricoides and Trichuris
▪ Internal Autoinfection = Strongyloides stercoralis, Capillaria philippinensis
▪ External Autoinfection = Enterobius vermicularis
▪ Larva of nematodes capable of heart and lung migration and can be found in
sputum specimen = Ascaris lumbricoides; Strongyloides stercoralis,
Hookworms
▪ Migration of large numbers of larvae through the lungs can cause Ascaris
pneumonitis or Loeffler´s syndrome characterized by bilateral, diffuse,
mottled pulmonary infiltrates and mild bronchitis associated with
peripheral eosinophilia. (p1264, Henry 23rd ed)
▪ Ground itch or creeping eruption is caused by larval penetration of
hookworms through the skin
▪ Parasites that can be recovered from the sputum
Larva of Ascaris lumbricoides; Strongyloides stercoralis, Hookworms
Paragonimus ova, Echinococcus granulosis hooklets from hydatid cysts,
Entamoeba histolytica trophozoites, Cryptosporidium parvum oocysts,
Entamoeba gingivalis, Trichomonas tenax
▪ Ascaris eggs who do not have the mamillated coat = decorticated e
▪ Ova is barrel shaped with prominent bipolar plugs = Trichuris trichiura
▪ Commonly known as threadworm = tron yloides stercoralis
▪ Capillaria philippinensis = eggs closely resembles Trichuris trichiura
▪ Trichinella spiralis = can be killed by cooking meat at 77°C (177°F) or by
freezing meat at -15°C for 20 days or -30°C for 6 days
▪ Trichinella spiralis adults are found in the M N N
▪ Loa loa = African eye worm
▪ Sheathed microfilariae = Wuchereria bancrofti, Brugia malayi, and Loa loa
▪ Unsheathed microfilariae = Onchocerca volvulus
▪ Onchocerca volvulus = Convoluted worm; causes river blindness;
▪ Dracunculus medinensis = fiery serpent of the Israelites, guinea worm
d. Cestodes/Tapeworms
▪ Parts of Tapeworm Adult
o Head = SCOLEX
o Region of growth = NECK
o Body of tapeworm, consists of proglottids (segments)= STROBILA
▪ Stain used to aid in counting the lateral uterine branches of Taenia = N N
▪ Eggs with hexacanth embryo and radially striated shells = N
▪ Has 7-15 uterine branches; causes neurocysticercosis = Taenia solium
▪ Third species of Taenia = Taiwan Taenia, also known as Taenia asiatica
▪ Taiwan Taenia infects domesticated pigs and wild boars.
▪ Adult Taenia are irritated by alcohol. Passage of proglottids results after a
drinking bout
▪ Double-pored tapeworm (dog tapeworm) = Dipylidium caninum
▪ Broad or Fish Tapeworm; Spoon-shaped or spatulate scolex, rosette uterus;
cestode with two intermediate hosts (copepods and fresh water fish); cestode
with operculated eggs = M M
▪ Cestode that resembles the adults of Diphyllobothrium latum = M
▪ Scolex with rostellum and crown of double hooks = N M
▪ Dwarf tapeworm, eggs with bipolar filaments = M N N N
▪ Definitive host of Echinococcus granulosus =
▪ Stage of Echinococcus granulosus found in the intermediate host =
C
23
 ▪Criteria for Cure: recovery of the scolex; negative stool exam for 3 months ▪ Entamoeba histolytica = pathogenic Entamoeba, causes colonic
after treatment perforation, amoebic liver abscess
e. Trematodes ▪ Free-living amoeba associated with keratitis and granulomatous amoebic
▪ Trematodes have two intermediate host, except Schistosomes encephalitis = C N M
o 1st Intermediate Host = SNAIL . Largest protozoan and the only ciliate capable of infecting humans id ey
o 2 Intermediate Host
nd e cr uc eus N MC
- Paragonimus westermani = freshwater crab . Flagellates
- Clonorchis, Opistorchis and Heterophyes = fish ▪ Flagellate that causes the ping-pong infection; can be demonstrated in the
- Fasciola, Fasciolopsis = aquatic vegetation
urine, vaginal and urethral discharges; has characteristic jerky tumbling
- Echinostoma = snail
▪ Characteristics of Schistosomes motion = C M N N
o Infective stage is cercaria; non-hermaphroditic, has separate sexes ▪ Trichomonas vaginalis has an undulating membrane that extends only
o One intermediate host: snail half the length of the body and is isolated in vaginal fluid, prostatic fluids
o Eggs are non-operculated; have spines or knob or sediments of freshly passed urine.
o Cercaria minus the tail = H ▪ Pentatrichomonas hominis (previously called Trichomonas hominis) is
o Causes swimmer’s itch = SCHISTOSOMA JAPONICUM characterized by 3-5 anterior and one posterior flagella; a prominent rod-
▪ Morphologic Appearance of Trematodes like object known as axostyle that protrudes from the posterior end; and an
o Eggs with small lateral spine or knob = Schistosoma japonicum undulating membrane extending length of the body of the parasite.
o Eggs with large lateral spine = Schistosoma mansoni ▪ Infective stage of Leishmania spp in humans: PROMASTIGOTE
o Eggs with large terminal spine = Schistosoma haematobium ▪ Giardia lamblia
o Eggs with old fashioned light bulb appearance = Clonorchis sinensis o Falling leaf motility; “Old man with eye glasses” trophozoite
o Eggs with abopercular thickening = Paragonimus westermani o Gay Bowel Syndrome and steatorrhea
▪ Mode of transmission of C. sinensis = ingestion of metacercaria in fish o Specimen: duodenal aspirates using Entero-String test
▪ Trematode eggs that hatches only after the egg is ingested by the first i. Toxoplasmosis
intermediate host: Clonorchis sinensis, Opistorchis spp ▪ Definitive host of Toxoplasma gondii = C
Heterophyid flukes (Heterophyes, Metagonimus, Haplorchis) j. Plasmodium sp.
▪ 90% of malaria cases are caused by . ci ru d . i
▪ Trematodes that release fully embryonated eggs in the stool ▪ Black water fever and cerebral malaria = Plasmodium falciparum
Clonorchis sinensis, Opistorchis sp ▪ Definitive host MOSQUITO Intermediate host: HUMANS
▪ Trematode associated with cholangiocarcinoma: Clonorchis sinensis ▪ Infective stage to humans =
f. Amoeba Infective stage to mosquitoes = M C
▪ Motile, feeding stage of amoeba = TROPHOZOITES ▪ Malarial species associated with malarial relapse = . i d . e
▪ Entamoeba histolytica vs Entamoeba hartmanni = size ▪ Crescent-shaped gametocytes; associated with double ring forms and
▪ Entamoeba histolytica vs Entamoeba dispar = molecular test applique forms; only ring and gametocyte forms are found in the blood film
▪ Amoeba without cyst stage: Entamoeba gingivalis PLASMODIUM FALCIPARUM
▪ Band forms = P. malariae

24
 ▪ Schuffner’s dots, enlarged RBCs = P. vivax . Concentration Techniques
▪ Gametocyte Male gametocyte = MICROGAMETOCYTE ▪ Zinc Sulfate Flotation Technique
Female gametocyte = MACROGAMETOCYTE o Specific gravity of Zinc sulfate = 1.180
▪ Rapid Diagnostic Tests detect for HRP-2 (histidine-rich protein 2) or p-LDH o Operculated eggs and eggs with spine are not recovered
▪ Knott’s Concentration Technique =
(parasitic-lactate dehydrogenase)
▪ Formalin-Ether Concentration Technique
HRP-2 = for Plasmodium falciparum o Ethyl acetate is used as a substitute for ether
p-LDH = for all four Plasmodium species Top layer – ethyl acetate 3rd layer – layer of formalin
k. Ectoparasites and Vectors 2nd layer – plug of fecal debris 4th layer – parasitic elements
▪ Phthirus pubis (genital/pubic louse, an ectoparasite) = CRAB LOUSE q. Additional Techniques
▪ Can serve as vector for Ascaris and Trichuris = COCKROACH ▪ Sheather’s sugar flotation technique = CRYPTOSPORIDIUM
▪ Vector for Wuchereria bancrofti = Anopheles, Aedes, Culex mosquito ▪ Harada-Mori = Strongyloides and Hookworms
▪ Vector for Brugia malayi = Mansonia mosquito ▪ Baermann funnel Technique = Strongyloides
▪ Primary Vector for malaria = Anopheles flavirostris ▪ Cellophane Tape (Scotch tape) preparation = Enterobius vermicularis
▪ Vector for Trypanosoma brucei complex = Tsetse fly (Glossina sp) <23.> Mycology
▪ Vector for Leishmania = sandfly (Phlebotomus sp) . Fungal structures and infections
l. Specimen ▪ Mass of hyphae, branching and intertwining structure of molds = Mycelia
▪ Paragonimus = Sputum/stool ▪ Microsporum = skin and hair
▪ Onchocerca volvulus = Skin snips ▪ Epidermophyton = skin and nails
▪ Trichinella spiralis = Muscle biopsy ▪ Trichophyton = skin, nails and hair
▪ Filaria, Plasmodia, Trypanosomes, Babesia = Blood ▪ Produces ascospores = Saccharomyces
m. Preservation of stool . Microscopic identification
▪ the stool must be REFRIGERATED if not analyzed within 1 hour ▪ 10 or 20% KOH = diss es er i
▪ Stool Preservatives ▪ Lactophenol cotton blue = lactic acid, phenol, and cotton blue kills, preserves
o 10% formalin o merthiolate-iodine-formalin and stains cell walls of fungi
o Schaudinn’s fluid o sodium acetate-formalin ▪ Used for sharp delineation of fungal elements in clinical specimens by
o polyvinyl alcohol fluorescence microscopy = CALCOFLUOR WHITE STAIN
n. Kato-Thick and Kato-Katz Technique ▪ Periodic acid Schiff = stains chitin in the cell wall of fungi
▪ Kato-Katz uses measured amount of stool and is used for egg counting; Kato- ▪ India Ink or nigrosin = demonstrates capsules of Cryptococcus
Thick does not use measured amount of stool c. Biochemical Tests
▪ Materials: cellophane, glycerin/glycerol, malachite green ▪ Germ tube test
o. Staining Technique For identification of C N C N
▪ Iodine mount = destroys trophozoites Reagent: rabbit plasma or serum; or fetal calf serum
▪ Modified Acid Fast Stain = for Cryptosporidium, Cyclospora, Isospora Incubate inoculum and reagent at 35°C for 2.5 to 3 hours
▪ Urease test = for Cryptococcus neoformans

25
d. Culture Media <24.> Virology
▪ Medium used to stimulate chlamydospore production of Candida albicans = a. RNA Viruses
CORNMEAL AGAR ▪ Arenavirus, Bunyavirus and Calicivirus (Norwalk)
▪ Stain in Aman medium = COTTON BLUE
▪ Coronavirus (Coronavirus, SARS)
▪ Czapek’s medium is used for isolation of = Aspergillus
▪ Bird seed/ Niger seed agar/ Staib medium detects for production of the ▪ Filovirus (Marburg, Ebola)
enzyme phenol oxidase. It is used for identification of Cryptococcus ▪ Flavivirus (Yellow fever, Dengue)
neoformans where it produces black-brown colonies. ▪ Orthomyxovirus (Influenza viruses)
▪ Rice agar is used to differentiate Microsporum canis from Microsporum ▪ Paramyxovirus (mumps, measles, Respiratory Syncytial virus)
audouinii ▪ Picornavirus (Poliovirus, Coxsackie, Echovirus, HepA virus, Rhinovirus)
▪ 2 antibiotics added in the Saboraud’s dextrose agar in order to inhibit ▪ Reovirus (Reovirus and Rotavirus)
bacteria and saprophytic fungi = Cycloheximide and Chloramphenicol
▪ Rhabdovirus (Rabies virus)
e. Fungi and their Characteristics
▪ Hair baiting test – positive result is V-shaped penetration; used to ▪ Togavirus (rubella; Eastern, Western & Venezuelan encephalitis)
differentiate Trichophyton mentagrophytes from Trichophyton rubrum ▪ Retrovirus (HIV, HTLV 1 and 2 virus)
▪ Characterized by tear-shaped microconidia; shows wine-red pigment in b. DNA Viruses
culture medium and gives a negative hair baiting test result = Trichophyton ▪ Adenovirus
rubrum ▪ Herpesvirus (HSV 1 and 2, VZV, EBV, CMV)
▪ Epidermophyton floccosum = Large, multicelled, club-shaped, smooth- ▪ Hepadnavirus (Hepatitis B virus)
walled macroconidia. Single or in clusters of two to three; microconidia
▪ Papovavirus (Papilloma and Polyoma virus)
not formed
▪ Chromoblastomycosis is often associated with the presence sclerotic ▪ Parvovirus (Parvovirus B19)
cells. Sclerotic cells, also known as Medlar bodies, are globe-shaped, cigar- ▪ Poxvirus (Variola)
colored thick-walled structures that are described as copper pennies. c. Characteristics of Viruses
▪ Dermatophytes that fluoresce under Wood’s lamp: ▪ The largest DNA virus: poxvirus
Microsporum canis Microsporum audouinii ▪ The smallest DNA virus parvoviridae
▪ Spaghetti and Meatballs appearance and the agent of Tinea versicolor
▪ The largest RNA virus: paramyxoviridae
Malassezia furfur
▪ Spherules with endospores = Coccidioides immitis ▪ The smallest RNA virus: enteroviruses (Picornaviridae)
▪ Broad-based budding yeast cells = Blastomyces dermatitidis ▪ The only single stranded DNA virus parvovirus
▪ Mariner’s wheel appearance = Paracoccidioides brasiliensis ▪ The only double stranded RNA virus reovirus
▪ Rose gardener’s disease = Sporothrix schenckii d. Steps in Viral Replication
▪ Fungi with aseptate hypha = Mucor, Absidia, Rhizopus ▪ Attachment/Adsorption, Penetration, Uncoating, Eclipse stage, Assembly
▪ Serologic tests for Histoplasmosis detect for the H and M band. Patients and Release
with Histoplasma capsulatum infection may demonstrate positive reactions
▪ 1st step = Adsorption/Attachment and Penetration
with serologic tests for Blastomyces dermatitidis since their antigens are
similar ▪ In enveloped viruses (i.e. influenza virus) the cytoplasmic or nuclear
membrane surrounds the nucleocapsid to form an envelope.
e. Cytomegalovirus is best isolated using Human embryonic fibroblasts

26
 . Swab for virus collection
▪ cotton, rayon and dacron on plastic shafts
▪ do not use calcium alginate since it can bind and inactivate virus
▪ do not use wooden shaft since wood and charcoal is toxic to viruses
. Storage Temperature of Viral specimen
▪ short term delay = refrigerate at de C
▪ long term delay = snap frozen at de C
. Viral Inclusion bodies
▪ Councilman bodies/ Torres bodies – yello e er irus
▪ Negri bodies – rabies virus
▪ Owl’s eye inclusion – cytomegalovirus
▪ Cowdry bodies – HSV, VZV, Polio
▪ Guarnieri bodies – small pox virus
i. Hexon is a group reactive antigen found in all adenovirus types except those of
avian origin
. Adenovirus-induced cytopathogenic effect = rounding & clumping (grape-like
cluster)
. Varicella zoster virus causes chicken pox and shingles.
. Human Papillomavirus (HPV) causes genital warts and can be a predisposing
factor to cervical carcinoma.
. Koplik’s spots are associated with = MEASLES
. Picornaviruses
▪ Ether-resistant viruses = no envelope; i.e. picornavirus (e.g. rhinovirus)
▪ Acid sensitive = rhinovirus; Acid resistant = enteroviruses
. Rabies virus is highly pathogenic and causes acute central nervous system
disease of man and animals
. Serological tests for viruses uses paired sera:
Acute-phase specimen - collected when clinical signs first appear
Convalescent-phase specimen - Collected after ee s
. SMALLPOX is the virus that has been eradicated worldwide
r. Slapped cheek appearance and aplastic anemia are associated with
s. Rotavirus causes acute viral gastroenteritis especially in children during
winter months
. Mad cow disease, spongiform encephalitis, kuru & scrapie: caused by PRIONS
CLINICAL MICROSCOPY
<1.> Urine
. Formation
▪ Normal daily urine output = d y
▪ urine output = polyuria urine output = oli uria
▪ Major inorganic substance of urine = C loride
▪ Organic substances in urine: urea (major), creatinine, uric acid
▪ Day to night urine ratio =
▪ Sequence of Urine formation: Afferent arteriole → Glomerulus →
Bowman’s space → Proximal convoluted tubule → Descending Loop of Henle
→ Ascending Loop of Henle → Distal convoluted tubule → Collecting Duct
▪ The renal artery supplies blood to the kidney. The human kidneys receive
approximately 20-25% of the blood pumped through the heart at all times.
▪ Rate of renal blood flow = m /day
▪ Filtering apparatus of nephrons = lomerulus
▪ Specific gravity of filtrate from the glomerulus = .
Size of molecules that can pass through the glomerulus = altons
▪ Responsible for majority of the reabsorption; reabsorbs glucose, amino acids,
water and electrolytes = ro imal Con oluted tubules
▪ Site of cast formation, target of ADH (vasopressin) = C and C
. Renal Function Test
▪ Inulin clearance test, creatinine clearance test, cystatin C are tests for:
lomerular iltration
▪ Gold standard for glomerular filtration = inulin clearance test
▪ Greatest source of error in clearance test = improperly collected urine
sample
▪ Specific gravity and osmolarity, Mosenthal and Fishberg concentration tests
are tests for tubular reabsorption/concentration
▪ Freezing point depression osmometers are most commonly used to
measure osmolality
▪ PAH, PSP, urinary ammonia, titratable acidity, urine pH are tests for
TUBULAR SECRETION
▪ Unit for microalbumin: ug/min
27
c. Specimen . Foam formation
▪ Volume capacity of container for routine urine analysis = 50mL ▪ White foam = increased protein; Yellow foam = bilirubin p ena opyridine
▪ Recommended volume for routine urine analysis = 10-15mL (12mL) . Odor of urine
▪ First morning urine specimen is recommended because: i is re ▪ Mousy/musty odor = N N
c ce r ed ▪ Sweaty feet urine odor = C C M
▪ A pregnant woman may test negative for urine hCG due to a possible diluted ▪ Rotting fish urine odor = M MN
specimen. ▪ Sweet/Fruity urine odor = N
▪ Changes in an unpreserved urine ▪ Cabbage odor = M NN M N
o Increased: pH, bacteria, odor, nitrite . Specific Gravity
o Decreased: glucose, ketones, bilirubin, urobilinogen ▪ In Diabetes mellitus: urine volume is and urine SG is
▪ Proper container for urine in urobilinogen determination = AMBER ▪ In Diabetes insipidus: urine volume is and urine SG is
▪ Urobilinogen can be determined using an early afternoon specimen ▪ Proper use of urinometer (hydrometer)
▪ Urine specimen type used in analytes with diurnal variation = TIMED o Adequate amount of urine is poured into a proper sized container
SPECIMEN o Urinometer is added with a spinning motion
▪ Chain of Custody refers to the step by step documentation of handling and o Reading is taken at the bottom of the urine meniscus
testing of legal specimens ▪ SG above 1.010 = N
▪ DONOR refers to the individual who submits a urine sample for drug testing SG fixed at 1.010 = N
(Strasinger) ▪ For every 3°C above or below the calibration temperature of the urinometer,
▪ Purpose of bluing agent in drug test = to prevent adulteration of specimen one should add or subtract 0.001 to the specific gravity to correct for the
▪ Requirements for Drug testing: Temperature . . de C temperature.
Volume m ▪ Refractometer or TS (total solids) meter determines specific gravity based
d. Color and Appearance on refractive index.
▪ Reddish-orange color of urine/body fluids = ri ampin ▪ Calibrating Solution for refractometer:
▪ Portwine red urine = porp yrins Distilled H2O = . 5% NaCl = . 9% sucrose = .
▪ Smoky red brown urine = ematuria i. pH
▪ Color imparted by carotene in urine = oran e ▪ vomiting, vegetarian diet, renal tubular acidosis = N N
▪ Color imparted by atabrine (antimalarial drug) in urine = yello ▪ high protein diet, DM, cranberry juice = C C N
▪ Condition associated with the presence of oily looking substance on top of . Chemical Examination
urine = nep rotic syndrome ▪ Most important chemical parameter in the identification of crystals = p
di. Clarity ▪ Parameter with the longest reading time = C
▪ In assessing urine color and clarity, a well-mixed specimen must be used, ▪ Detected in the 11 pad of the reagent strip = C
th C C
and the urine must be viewed through a clear container. ▪ Parameters affected by ascorbic acid N C
▪ Grading N and C
o Print is blurred through the urine = cloudy ▪ Parameter in the reagent strip with purple positive result = KETONES AND
o Few particulates, print easily seen through the urine = a y LEUKOCYTE ESTERASE

28
 er e ic es s r uri e
▪ Positive nitrite on reagent strip testing may indicate the presence of gram Sulkowitch test = ciu
negative bacilli Fantus test = ride
▪ Positive color of glucose pad in reagent strip if chromogen is potassium Heller’s test = r ei
iodide (multistix) = N N Rothera’s test = e es
▪ Principle of Clinitest: copper reduction (based on ability of sugars to Legal’s test = e es
reduce cupric into cuprous ion. Seliwanoff test = ruc se
▪ Sensitivity range of glucose parameter of reagent strip = m /d Rubner’s test = c se
▪ Sensitivity range of bilirubin parameter of reagent strip = . . m /d Obermayer test dic
▪ Sensitivity range of Ictotest tablet test for bilirubin = . . m /d Blondheim’s test e i y i
▪ Ehrlich units is the unit used in reporting = N N Sodium-Nitroprusside e es
▪ Of the routine chemical tests performed on urine, the most indicative of Cyanide-Nitroprusside cys i e
renal disease is the PROTEIN. Silver-Nitroprusside cys ei e
▪ Results: 3% SSA positive and negative protein rgt strip = N In Mucopolysaccharidosis: Reagent Positive result = ie
N MN
. Microscopic Examination
▪ SSA result
▪ The light source in the microscope is equipped with RHEOSTAT, to regulate
Noticeable turbidity (6-30mg/dL) = TRACE
the intensity of the light.
Turbidity with granulation but no flocculation (100-200mg/dL) =
▪ Phase contrast microscope is used for sediments with low refractive
Turbidity with granulation and flocculation (200-400mg/dL) =
index
▪ Positive parameter in starvation/vomiting = KETONES
▪ Hoffman (modulation contrast) and Nomarski (differential interference
▪ Most predominant ketone body = C C
contrast) are types of interference contrast microscope
▪ Ketone body that is directly detected by sodium nitroprusside =
▪ Polarizing microscope is preferred for oval fat bodies, lipids and crystals.
C C C C
▪ Acetic acid enhances the nuclei of WBCs and lyses red blood cells. Acetic acid
▪ Principle of protein pad in reagent strip = PROTEIN ERROR OF INDICATORS
can also be used to dissolve amorphous phosphates. Warming of urine
▪ Protein error of indicator: protein acts as proton acceptor; indicator acts
dissolves the amorphous urates.
as proton donor
▪ Yeast cells are often mistaken as RBCs during urine microscopic
▪ Speckled pattern in the blood pad of urine reagent strip suggests
examination. They can be differentiated using acetic acid. Acetic acid can
HEMATURIA.
lyse the red blood cells but not the yeast cells.
▪ In the presence of free hemoglobin/myoglobin, uniform green-blue color
▪ Normal value of hyaline casts in Addis count =
is produced.
▪ WBCs and RBCs are reported in HPF
▪ Chemical parameter that serves as screening test for UTI = N d
▪ RBC in hypotonic urine = GHOST CELLS
▪ Principle of the automated dipstick readers: reflectance ▪ WBCs in hypotonic urine = GLITTER CELLS
▪ Eosinophils are stained using N N. A finding of is
spectrophotometry
considered to be clinically significant. Increased eosinophils are associated
▪ Reagents strips must be stored in cool dry place
with acute/drug-induced interstitial nephritis
▪ Largest cell in the urinary sediment = Squamous epithelial cells

29
▪ Clue cells are indicative of bacterial vaginosis caused by Gardnerella vaginalis ▪ Renal failure casts = C . Most commonly seen broad casts are
▪ Transitional epithelial cells might be increased in the urine following an invasive GRANULAR and WAXY.
urological procedure such as catheterization. They are estimated as rare, few, ▪ Casts disintegrate in dilute alkaline urine.
moderate or many per high power field. ▪ Casts are examined under subdued (low) light, enumerated using low
▪ RTE cells are larger than WBCs, more polyhedral and has eccentric nucleus power fields and scanned at the edges of the coverslip.
▪ Oval fat bodies are found in patients with NEPHROTIC SYNDROME ▪ Casts with one tapered end; formed at the junction of the ascending loop of
▪ Bubble cells are RTE cells that are characterized by large, non-lipid-containing Henle ad DCT, has the same significance as casts = C N
vacuoles ▪ Simultaneous occurrence of the findings of nephrotic syndrome and
▪ Approximately 75% of the renal calculi are composed of calcium oxalate or glomerulonephritis on the same specimen = C M N
phosphate. l. Clinical Significance
▪ Crystal that can assume the widest variety of shapes; can be lemon-shaped, ▪ Acute tubular necrosis is caused by heavy metals, drugs, hemoglobin and
barrel-shaped, rhombic, four sided plates, hexagonal, wedges, rosettes = C myoglobin, viral infections, pyelonephritis, allergic reactions malignant
C infiltrations, salicylate toxicity and transplant rejection
▪ Majority of the renal calculi contains = C C M ▪ Ingestion of bananas, chocolate, walnuts, plums or certain medications can
H H cause false elevations of 5-HIAA.
▪ Crystal seen in Fanconi’s syndrome = C N C ▪ GOODPASTURE SYNDROME is an autoimmune condition characterized by
▪ Crystal seen in ethylene glycol (anti-freeze agent) poisoning = calcium oxalate presence of anti-glomerular basement membrane antibodies and anti-
monohydrate (dumbbell shaped, elongated hourglass appearance) alveolar basement membrane antibodies.
▪ Crystals found in patients with liver disease = N CN N ▪ Presence of ANCA (anti-neutrophilic cytoplasmic antibodies) = WEGENER’S
▪ Thorny apple crystals = MM N M C GRANULOMATOSIS
▪ Struvite crystals are also known as triple phosphate or ammonium magnesium ▪ ALPORT SYNDROME is a genetic disorder showing lamellated and thinning
phosphate. They are coffin lid shaped crystals found in alkaline urine. glomerular basement membrane
▪ Apatite crystals are also known as = C C M ▪ Alkaptonuria is an inherited defect associated with increased urinary
▪ Lignin test is used to determine the presence of sulfonamides in urine sample. excretion of homogentisic acid. This is due to deficiency of enzyme
One drop of urine is placed on a newspaper, after which one drop of 10% HCl is homogentisic acid oxidase. Urine with homogentisic acid has the tendency
added. to turn dark brown to black upon standing
▪ Most common parasite encountered in the urine = C M N N ▪ Phenylketonuria is an inborn error of metabolism characterized by
▪ Parasite that is a fecal contaminant in the urine = N MC deficiency of the enzyme phenylalanine hydroxylase, which causes the
▪ Represents disintegration of the cellular casts = N C accumulation of phenylpyruvic acid in the urine, giving the urine its typical
▪ Final degenerative form of casts, characterized by brittle consistency and jagged mousy or musty urine odor.
ends = WAXY CAST ▪ NMP-22 (nuclear matrix protein) can be used as tumor marker for urinary
▪ Sequence of Cast development: bladder carcinoma.
hyaline >> cellular >> coarsely granular >> finely granular >> waxy

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<2.> Semen o. Sperm count
. Spermatogenesis: ▪ Diluting fluid: sodium bicarbonate + formalin
▪ Spermatogonium is the youngest form Chilled water
▪ Spermatogonia → primary spermatocytes → secondary spermatocytes→ ▪ Dilution: 1:20
spermatids . Computer Assisted Semen Analysis (CASA) provides objective determination
▪ Spermatids then mature into spermatozoa, also known as sperm cells. of sperm velocity, sperm trajectory, sperm concentration and sperm
. ACROSOMAL CAP should encompass one-half of the head of the sperm cell and morphology.
should cover approximately two-thirds of the nucleus. It contains enzymes that . Grading of Sperm Motility
are critical for ovum penetration. ▪ 4.0 = rapid, straight-line motility
c. Thickest part of the tail surrounded by mitochondrial sheath M C ▪ 3.0 = slower speed, some lateral movement
d. Seminal vesicle is the major contributor to seminal fluid volume, accounting to ▪ 2.0 = slow forward progression, noticeable lateral movement
approximately 60% of the total semen. It is responsible for the fructose content ▪ 1.0 = no forward progression
of the semen. ▪ 0 = no movement
e. Prostatic fluid contains zinc, acid phosphatase, proteolytic enzymes and <3.> Cerebrospinal fluid
citric acid. r. CSF is collected through lumbar puncture between the 3rd and 4th or the 4th
. The most common cause of male infertility, characterized by the hardening of and 5th lumbar vertebrae.
veins that drain the testes, which causes the blood from the adrenal vein to s. If three tubes are collected for CSF, the first tube is sent to the chemistry or
flow into the spermatic vein = VARICOCELE serology department, the second tube is sent to the microbiology department
. Specimens for seminal fluid analysis are collected following a period of sexual while the last tube is sent to the hematology section for the cell count.
abstinence of from 2 to 3 days to not longer than 5 days. Specimens collected . If there is only a single specimen for CSF analysis, the specimen must be
following prolonged abstinence tend to have higher volumes and decreased delivered to the microbiology section first, then to hematology for cell count and
motility finally to chemistry and serology
. Three glass collection method: the second specimen= serves as control for u. An estimated 70% of CSF is derived by ultrafiltration and secretion through the
bladder and kidney infection choroid plexuses. The ventricular ependymal lining and the cerebral
i. Stain used to assess sperm morphology = H subarachnoid space account for the remainder. – p481, Henry 22nd ed.
. Stain used to assess sperm viability = NN N . Uneven blood distribution in 3 tubes, clear supernatant fluid, clot formation, no
. Neutral alpha-glucosidase assesses the function of M erythrophagocytes/erythrophages = M C
. If seminal fluid specimen for fructose could not be analyzed within two hours, the . Even blood distribution in 3 tubes, xanthochromia, no clot formation, presence
specimen must be N of erythrophagocytes = N C N M
. Enzymes that can liquefy semen = CHYMOTRYPSIN . Pellicle formation in CSF after 12-24 hour incubation at refrigerated
. Florence test is a simple screening test for the presence of semen. It detects for temperature indicates the possibility of tuberculous meningitis.
choline, using the Florence reagent which is made up of potassium iodide, y. Normal adult CSF ranges between 0-5 WBCs per uL.
iodine crystals and distilled water. . Increased number of WBCs is CSF = PLEOCYTOSIS

31
 . Lymphocytes are predominant in CSF of adults. In neonates, however, <6.> Feces/Stool
Monocytes predominate. . Urobilinogen is synthesized in the intestines from bilirubin diglucoronide (B2).
. Cloudy CSF must be diluted . Color of stool in Iron ingestion (in vitamins), charcoal, Bismuth, upper GIT
. Normal Value of protein in CSF = m /d bleeding, melena =
. Laboratory tests for CSF protein analysis = TURBIDIMETRIC, DYE-BINDING c. Color of stool in Barium sulfate ingestion, bile duct obstruction =
. OLIGOCLONAL BANDS – suggestive of multiple sclerosis, can be found in d. Fecal Occult Blood Test (FOBT) = detects “ idden blood”; used as a screening
Guillain-Barre syndrome, bacterial meningitis, viral encephalitis, subacute test for C C C NC
sclerosing panencephalitis (SSPE), and neurosyphilis e. Guaiac test is the preferred method for fecal occult blood test. Positive result is
. CSF glutamine is elevated in the CSF of patients with Reye’s syndrome and the formation of blue color.
hepatic coma. . The myoglobin and hemoglobin of ingested meat and fish have peroxidase
<4.> Serous fluids activity that may falsely indicate the presence of occult blood. Bacteria in the
. Accumulation of fluid in between the visceral and parietal membrane = N bowel, as well as ingested vegetables and fruits, such as horseradish, turnips,
. Conditions associated with Transudates es i e e r i ure e r ic bananas, black grapes, pears, and plums, have peroxidase and can falsely elevate
sy dr e irr sis fecal peroxidase activity. False-negative tests occur in the presence of large
c. Conditions associated with Exudates ec i i cy i amounts of vitamin C and other antioxidants.
d. Spontaneous clot formation usually occurs with exudates but not with . APT test is used to differentiate maternal blood (HbA) from fetal blood (HbF).
transudates. (p223, Strasinger 5th ed) Reagent for the APT test is 1% NaOH
e. Psammoma bodies in peritoneal fluid that contains concentric striations of . Normal pH or the stool = . .
collagen-like material can be seen in benign conditions, ovarian malignancies and i. As few as t ree fecal leukocytes may indicate invasive infection
thyroid malignancies. . Conditions that can lead to appearance of fecal WBCs
. Normal peritoneal fluid appears CLEAR, PALE YELLOW Yersinia, EIEC, Campylobacter, Salmonella, Shigella
. Intestinal perforation is associated with high levels of alkaline phosphatase. k. Conditions that will not lead to appearance of fecal WBCs
. Specimen of choice for assessment of intra-abdominal bleeding = eri e e Staphylococcus, Vibrio, parasites, virus
l. Quantitative fecal fat analysis is used as a confirmatory test for steatorrhea.
<5.> Synovial Fluid As discussed, quantitative fecal analysis requires the collection of at least a 3-
. Normal volume of synovial fluid = . day specimen. Van de Kamer titration serves as the gold standard.
. Because synovial fluid is an ultrafiltrate of plasma, chemistry test values are
approximately the same as serum values. <7.> Gastric Analysis
c. Normal synovial fluid glucose should not be more than 10mg/dL lower than the a. Gastric specimen can be obtained via gastric brushing, gastric lavage, as well
blood value as induced-vomiting.
d. Neutrophils with precipitated rheumatoid factor = r cy es ce b. Gastric Tubes
e. DO NOT USE powdered anticoagulant – since it can interfere with crystal ▪ Inserted through the mouth = H
identification ▪ Inserted through the nose =
. DO NOT USE acetic acid when diluting synovial fluid for WBC count. Acetic acid can
cause clotting of synovial fluid. Use hypotonic saline (0.3%) instead, to lyse RBCs

32
 \ -

Wala sa notes
c. Diagnex blue test is used in the tubeless gastric analysis. The specimen in this
test is urine.
d. BAO = Basal Acid Output
HEMATOLOGY
MAO = Maximal Acid Output
e. Gastric Stimulants Test Meals 1. Skin puncture is used for:
▪ Most preferred gastric stimulant = __________________________ a. Infants less than 6 months of age
▪ Used to assess success of vagotomy = __________________________ b. Young children if only small amount of blood is needed
f. Diseases c. Adults with poor veins, or when veins cannot be used because of IV infusions, burns,
▪ Zollinger Ellison Syndrome = elevated
extreme obesity, thrombotic tendencies, and with point- of-care testing or with patients
▪ Pernicious Anemia = anti-parietal cell antibodies
<8.> Respiratory Tract Specimen performing tests at home
. Bronchial washings and broncoalveolar lavage (BAL) are respiratory tract 2. Site of Skin Puncture
specimens that can be used for cytology. a. <1 yr of age = medial or lateral portion of the plantar surface of the foot
. Sputum crystals that represent eosinophil degradation = H b. >1 yr of age = Second, third or fourth finger (middle finger is the most commonly used);
puncture should be perpendicular to the fingerprint on the palmar surface of the end
c. Elements found in patients with bronchial asthma = Creola bodies, Curschmann portion of the finger, slightly off-center
spirals, Charcot Leyden crystals
c. Sites used for obtaining arterialized capillary blood = FINGER, EARLOBES, HEEL
d. Anchovy or rusty red sputum = obar pneumonia treptococcus pneumoniae
3. Depth of Skin Puncture = not more than 2mm
e. Myelin globules in sputum have no clinical significance but may be mistaken as
Blastomyces 4. Capillary blood is arterialized by warming the ear, finger or heel at 42°C before taking the
sample, for blood gas analysis.
<9.> Pregnancy Testing
a. hCG is a glycoprotein hormone similar to H H d H. They have \ A blood film taken from a capillary puncture has sometimes been referred to as the “poor
5.
similar alpha chains by different beta chains man’s aggregation study”. (Clinical Hematology: Principles, Procedures and Correlations by
b. Biologic Test Ascheim-Zondek = immature female, mice Steininger)
Frank-Berman = immature female rats 6. Venipuncture procedure: (1) patient interaction; (2) assemble supplies and equipment; (3)
Friedman = mature, virgin, female rabbit venipuncture; (4) specimen preparation
Galli-Mainini = Male frog/Male toad 7. Site of Venipuncture: antecubital fossa
Hogben = female toad
a. Basilic Vein is located on the inner portion of the arm.
Kupperman = female rat
<10.> Amniotic Fluid b. Cephalic Vein is located on the edge of the outer arm.
. Gestational age can be assessed using amniotic fluid creatinine. c. MEDIAN CUBITAL VEIN is the most preferred vein since it is well-anchored in tissue and
. Creatinine can differentiate urine from amniotic fluid. does not roll when punctured.
c. Dark green color of the amniotic fluid indicates presence of 8. The gauge number of needle is inversely proportional to the diameter of the bore (bore size)
d. Dark brown color of the amniotic fluid indicates presence of H 9. Needle used in vacutainer tube is DOUBLE ended
e. Tests for neural tube defects and acetylc olinesterase 10. Most commonly used gauge numbers for adult phlebotomy = 19, 20, 21, 22
. Tests to assess severity of HDN 11. Length of needle used in venipuncture = 1.0 to 1.5 inches
Oxyhemoglobin can interfere during the OD450, since it can absorb light at
12. Color of needle hub of gauge number 21 = GREEN
410nm.

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13. Length of tourniquet = 18-20 inches; Width of tourniquet = 1 inch 21. Skin puncture order
14. Tourniquet should not be applied for more than: 1 minute a. Blood gases
15. Distance of tourniquet from the puncture site = 3-4 inches b. Slides/smear
16. Angle of needle to skin c. EDTA microcollection tubes
a. Venipuncture = 15-30-degree angle d. Other microcollection tubes ie, green or gray
b. Brachial arterial puncture= 45-60 e. Serum microcollection tubes
c. Femoral arterial puncture = 90 22. Hematologic Tests and Specimen Type(s) used:
17. Number of time(s) a technologist or a student can stick a patient = 2x a. EOFT, Autohemolysis test (H.Spherocytosis), Acid Serum Test (PNH) = DEFIBRINATED
18. Patients with IV lines: IV line should be stopped for 2mins 5mL of blood should be collected BLOOD
and discarded b. Haptoglobin and Vitamin B12 determination = SERUM
19. Anticoagulants: c. Fibrinogen Quantitation = CITRATED-PPP
a. Binds calcium to form soluble complex = CITRATE d. Methemoglobin quantitation = HEPARINIZED WHOLE BLOOD (fr Alba)
b. Binds calcium to form insoluble complex = OXALATE 23. Cell counting/Hemocytometry
c. Inhibits coagulation by chelating calcium = EDTA a. RBC diluting fluids should be isotonic. Examples of RBC diluting fluids include:
d. The liquid tripotassium salt of EDTA is thought to cause a 2-3% decrease in the ▪ Hayem’s fluid
\ hematocrit due to slight shrinkage of red blood cells (p86, Hematology: Principles and ▪ Gower’s fluid
Procedure by Barbara Brown) ▪ Toisson’s fluid
e. Inhibits coagulation by inhibiting thrombin = HEPARIN. Heparin acts with cofactor ▪ Bethel’s fluid
antithrombin III ▪ Formol-Citrate (Dacie’s fluid)
f. Anticoagulant to blood ratio in coagulation = 1:9 ▪ NSS (0.85% NaCl)
g. Concentration of EDTA = 1.5mg per mL of blood ▪ 3.8% Na Citrate
h. Concentration of Heparin = 15-20U/mL of blood b. WBC diluting fluids should lyse RBCs. Examples of WBC diluting fluids include:
20. Order of draw for evacuated tubes and syringe (Citrate – Heparin – EDTA – Fluoride) ▪ 2-3% glacial acetic acid with Gentian violet
a. Blood culture tubes (SPS, yellow top) ▪ 1% hydrochloric acid
b. Na citrate (light blue top, black top) ▪ Turk’s solution
c. Serum tubes (red top) c. Platelet Count diluting fluids include:
d. Heparin tubes (green t op) ▪ Rees-Ecker Diluent (Tocantin’s method, for LIGHT microscopy)
e. EDTA tubes (lavender top) ▪ 1% ammonium oxalate (Brecker-Cronkite method, for PHASE CONTRAST
f. Glycolytic inhibitor tubes (gray top, fluoride-oxalate) microscopy)

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d. Counting Chambers: k. WBC Estimates:

▪ Dimensions of Neubauer counting chamber = 3mm x 3mm x 0.1mm 2-5 WBCs/hpf = 4,000-7,000 WBCs/ɥL
• The depth of the Neubauer counting chamber is 0.1mm or one-tenth mm. 4-6 WBCs/hpf = 7,000-10,000 WBCs/ɥL
• Smallest square in the Neubauer counting chamber has an area of 0.0025mm2 6-10 WBCs/hpf = 10,000-13,000 WBCs/ɥL
▪ Dimensions of Fuchs-Rosenthal counting chamber = 4 x 4 x 0.2 mm 10-20 WBCs/hpf = 13,000-18,000 WBCs/ɥL
24. Blood Film Preparation:
▪ Dimensions of Speirs Levy counting chamber = 2 x 5 x 0.2 mm
a. Size of drop of blood = 2-3mm
e. If the WBC count is markedly elevated as in some leukemias, in which it may be as high
b. Distance of blood from the end of the slide = 1cm
as 100 to 300 x 109/L, a 1:200 dilution is used. (Brown)
c. Directions in Counting cells: cross-sectional/crenellation, longitudinal/lengthwise,
f. If WBC count is below 1.0 x 109/L, differential count may be performed by counting 50
battlement method
WBCs instead of 100.
g. A 200-cell differential is indicated when there is an abnormal distribution of cell types,
\ Battlement method of reading the blood smear uses a pattern of consecutive fields
d.
beginning near the tail on a horizontal edge; count three consecutive horizontal edge
such as when there is: fields (moving away from the tail), count two fields toward the center of the smear,
• More than 10% eosinophils count two fields horizontally (moving away from the tail), count two fields vertically to
• More than 2% basophils the edge. (p103, Hematology: Principles and Procedure by Barbara Brown)
• More than 11% monocytes e. Factors that affect the thickness of blood film
• More lymphocytes than neutrophils (except in children). For a thicker film For a thinner film
Results are averaged. Notation is made on the report that 200 WBCs were counted. Pressure decrease increase
h. Normal platelet count should show approximately 7-25 (8-20) per field Angle increase decrease
\i. Estimates: Size of drop of blood increase decrease
▪ Factor for platelets = # of platelets per field x 20,000 Speed of spreading increase decrease
▪ Factor for WBCs = # of WBCs per field x 2,000 f. Brilliant cresyl blue and new methylene blue are examples of: Supravital stains. Heinz
j. Platelet Estimates: bodies and reticulocytes are not visible in Wright-Giemsa-stained smear. They are
markedly decreased 0-49,000/uL demonstrated using supravital stains.
moderately decreased 50,000-99,000/uL g. Examples of Romanowsky Stain include: Wright, Giemsa, Wrights-Giemsa, Leishman,
slightly decreased 100,000-149,000/uL Jenner, May-Gruwald, MacNeal’s
low normal 150,000-199,000/uL h. pH of the buffer in Wright-Giemsa stain = 6.4-6.8
Normal 200,000 – 400,000/uL
slightly increased 401,000 – 599,000/uL
moderately increased 600,000 – 800,000/uL
markedly increased >800,000/uL

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i. Causes of excessive blue and red color in blood films 27. Variations in RBC shape:
EXCESSIVELY RED STAIN EXCESSIVELY BLUE STAIN a. Horn-like or helmet like cell = KERATOCYTES
▪ Too acid buffer (below 6.4) ▪ Too alkaline buffer (above 6.8) b. Sometimes known as burr cells, found in patients with kidney disease or uremia =
▪ Too acid stain solution ▪ Too alkaline stain solution ECHINOCYTES
▪ Insufficient staining time ▪ Excessive staining time
▪ Excessive washing ▪ Inadequate washing c. Abetalipoproteinemia, McLeod phenotype = ACANTHOCYTES
▪ Very thin smears ▪ Very thick smears d. Myelofibrosis = DACROCYTES
▪ Contaminants like chlorine ▪ Old smear (dried plasma produces blue e. Thalassemia, Mexican hat cells = CODOCYTES
▪ Exposure of buffer or stain background)
f. Megaloblastic anemia = MACRO-OVALOCYTES
to acid fumes ▪ Protein abnormality
▪ Old stain in which ▪ Use of heparinized blood sample g. Rh null = STOMATOCYTES
methanol is oxidized to ▪ Very high leukocyte count with many blasts \ h. Bite cells are often seen during acute hemolytic episodes of patients with G6PD
formic acid ▪ Low hematocrit deficiency. The bites in these cells probably represent Heinz body craters. (Clinical
Hematology: Principles, Procedures and Correlations by Steininger)
j. Sudan Black B = old specimen can be used, positive in myeloid leukemia 28. WBC Morphologic Changes:
k.Myeloperoxidase = requires fresh specimen, positive in myeloid leukemia a. Crumpled tissue appearance of monocyte-macrophages, Glucocerebrosidase deficiency
l.TDT = positive for lymphoid leukemia = Gaucher disease (monocyte/macrophages are PAS positive)
m.Non-specific esterase = positive for monocytes b. Pince-nez, dumbbell shaped, bilobed nuclei, hyposegmentation = PELGER HUET
n.Specific esterase = positive for granulocytes c. golden brown granules; defective lysosomes = CHEDIAK HIGASHI
o.Tartrate-resistant acid phosphatase = positive in hairy cell leukemia d. remnants of lymphocytes; found in patients with CLL = SMUDGE CELLS
p.In the Leukocyte alkaline phosphatase score test (Kaplow count), 100 neutrophils are e. Found in patients with bacteremia or septicemia = TOXC GRANULATION,
counted and graded from 0 (negative) -4 (strongly positive). LAP score is decreased in VACUOLATIONS, DOHLE BODIES
CML/CGL and increased in Leukemoid reaction f. LE cell (Lupus Erythematosus cell) is a neutrophil that has phagocytized (engulfed) the
\
25. RBC Morphology denatured nuclear material or a nuclear body of another cell.
a. RBC size = 6-8um in diameter g. Russell bodies are eosinophilic, large, homogeneous immunoglobulin-containing
b. RBC shape = biconcave disc inclusions usually found in a plasma cell undergoing excessive synthesis of
c. Variation in the size = anisocytosis immunoglobulin. Multiple aggregates of Russell bodies create what are known as Mott
d. Parameter that assesses the degree of anisocytosis = RDW (red cell distribution width cells.
e. Variation in the shape = poikilocytosis h. Sezary cells are T cells found in patients mycosis fungoides. show the characteristic
26. RBC Inclusions convoluted to cerebriform nucleus with mildly condensed chromatin and inconspicuous
a. Feulgen positive, DNA fragment = Howell Jolly bodies nucleoli.
b. denatured hemoglobin, found in G6PD deficiency =Heinz bodies 29. Additional Cells:
c. lead poisoning, pyrimidine 5’-nucleotidase deficiency, aggregates of ribosomes = a. Osteoblasts are often mistaken as PLASMA CELLS
Basophilic stippling b. Osteoclasts are often mistaken as MEGAKARYOCYTES
d. Iron granules, Prussian blue positive = Pappenheimer bodies

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30. Grading: 31. Hematocrit

i. Hypochromia: a. Adam’s Microhematocrit (uses Capillary tube)
▪ Normochromic = 1/3 of cell diameter ▪ Length of capillary tube: 70-75mm; Bore Size: 1.0-1.2mm
▪ 1+ = area of central pallor is ½ of cell diameter ▪ Capillary tubes should be at least two-thirds filled with blood
▪ 2+ = area of pallor is two-thirds of cell diameter ▪ Speed of centrifugation: 10000-12000 x g for 5 minutes
▪ 3+ = area of pallor is three quarters ▪ The measurement should be performed within 10 minutes to avoid merging of the
▪ 4+ = thin rim of hemoglobin layers.
j. Sickle cells, Basophilic stippling, Pappenheimer bodies, Howell Jolly bodies = Grade only b. Wintrobe’s Macrohematocrit (Wintrobe tube)
as positive ▪ Length of wintrobe tube: 115mm (11.5cm)
k. Macrocytes ▪ Bore size: 3mm
▪ 1+ (slight) = approx.. 25% of macrocytic RBCs seen in hpf ▪ Speed of centrifugation: 2000-2300xg for 30 mins
▪ 2+ to 3+ (moderate) = approx.. 25-50% macrocytic RBCs per hpf c. LAYERS OF BLOOD AFTER CENTRIFUGATION (Top to bottom)
▪ 4+ (marked) = >50% macrocytic RBCs per hpf ▪ Fatty layer
l. Poikilocytosis ▪ Plasma
▪ Within normal limits = 0-2 ▪ Buffy coat
▪ 1+ = 3-10 ▪ Pack red cells
▪ 2+ = 10-20 32. Erythrocyte Sedimentation Rate:
▪ 3+ = 20-50 a. Westergren method
▪ 4+ = >50 ▪ Length 300mm
m. Rouleaux Formation (Hubbard) ▪ Graduation lines 200mm
▪ Slight = 1-2 RBC chains are found per thin microscopic field ▪ Internal Bore 2.65mm; External Bore 5.5mm
▪ Moderate = 3-4 RBC chains are found per thin microscopic field b. Macrocytes causes false INCREASE ESR
▪ Marked = >5 RBC chains are found per thin microscopic field c. Microcytes causes FALSE DEC ESR
n. Codocytes, Spherocytes, Ovalocytes, Stomatocytes d. Increased hemoglobin causes FALSE INCREASE ESR
▪ Within normal limits = 0-2 e. Anemia causes false INCREASE in ESR;
▪ 1+ = 2-10 f. Rouleaux formation INCREASES ESR
▪ 2+ = 10-20 g. Poikilocytes causes DECREASE ESR
▪ 3+ = 20-50 h. Over-anticoagulation of blood causes false DECREASE
▪ 4+ = >50 i. A tilt of 3° can cause errors up to 30%
j. Increase in temperature causes false INCREASE
k. Decrease in temperature causes false DECREASE
l. DISPETTE- disposable plastic tubes with safety plug made of nonwettable and
nonabsorbent material that provides automatic zeroing of the specimen

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33. Zeta Sedimentation Rate b. ACID ELUTION

a. uses a special instrument, zetafuge ▪ PRINCIPLE: HbF are resistant to acid elution
b. Normal value of ZSR (Males and Females) = 40-51% ▪ REAGENT: Erythrosin B; Ehrlich’s/Mayer’s hematoxylin
34. Erythrocyte Osmotic Fragility Test ▪ RESULTS: Deep pink RBCs = inc HbF; Pale ghost cells = dec HbF
a. Decreased EOFT – sickle cells, target cells, hypochromic cells, reticulocytes 38. Sucrose hemolysis test is used as a confirmatory test for paroxysmal nocturnal
\
b. Increased EOFT – spherocytes, senile/old cells hemoglobinuria when the sugar water test is positive. Specimen for sucrose hemolysis test is
35. Eosinophil count citrated whole blood. (p193-194, Hematology: Principles and Procedure by Barbara Brown)
a. Phyloxine or Phloxine B diluting fluid 39. Automation – Coulter Counter
▪ PHYLOXINE/PHLOXINE B- stains eosinophils red a. Principle: electrical resistance/impedance
▪ SODIUM CARBONATE and WATER - lyses WBCs b. Ohm’s Law states that: voltage = current x resistance
▪ PROPYLENE GLYCOL - lyses RBCS c. Isotonic solutions conduct electricity better than cells do
b. Pilot’s solution – same as Phyloxine/Phloxine B diluting fluid plus: HEPARIN to inhibit d. Number of pulses is directly proportional to the number of cells
leukocyte clumping e. Voltage size/ Height of pulse is directly proportional to the cell size
36. Solubility Test for Sickle cells f. Cell Sizes RBC count = >36fL
a. Blood is added to a buffered salt solution containing a reducing agent such as sodium Platelet count = size: 2-20fL
hydrosulfite (dithionite) and a detergent-based lysing agent (saponin). The saponin g. For WBC count = RBCs should be lysed using lysing agent or lytic agent
dissolves membrane lipids causing release of hemoglobin from RBCs and dithionite h. Lyse-resistant RBCs = causes false increase in WBC count
reduces the iron from the ferrous to ferric. Deoxygenated HbS polymerizes in solution,
i. Correct WBC count when >5 nucleated RBCs are counted
causing turbidity
b. Positive result is characterized by turbidity or when lines behind the tube are not j. Formula for Corrected WBC count = (WBC ct x 100) divided by (nucleated RBC + 100)
visible. k. MCH is not often used in the classification of anemia
c. Sources of Error: l. Histograms (for RBC and platelet count)
False Negative False Positive ▪ X-axis = SIZE
- Holding tube close to card - Increased globulins and ▪ Y-axis = RELATIVE NUMBER
- Use of 10x75mm test tube lipids ▪ From RBC Histogram, MCV and RDW can be derived
- Patients recently transfused with normal - Increased WBCs, RBCs ▪ From Platelet histogram, MPV and PDW can be derived
blood and Hgb
m. Cytogram or Scatterplot
- Infants younger than 6 mos. - Addition of more blood
to reagent ▪ X-axis = internal cell complexity (result of side scatter)
- Anemic patients (hgb <7g/dL)
▪ Y-axis = volume or size (result of forward scatter)
n. WBC 3-part differential Lymphocytes: 35-90fL
37. Tests for Hemoglobin F quantitation
Mononuclear cells: 90-160fL
a. ALKALINE DENATURATION
Granulocytes: 160-450fL
▪ PRINCIPLE: HbF are alkali-resistant
o. WBC 5-part differential: neutrophils, monocytes, lymphocytes, basophils, eosinophils
▪ METHODS: Betke's Modification (NaOH); Singer Method

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p. Rule of 3 43. Hemoglobin determination (Cyanmethemoglobin)

▪ RBC ct in millions x 3 = hemoglobin a. Reagents:
▪ hemoglobin x 3 = hematocrit ▪ Converts Hgb to methemoglobin = Potassium ferricyanide
q. RBC indices normal values: ▪ Converts methemoglobin to cyanmethemoglobin = Potassium cyanide
MCV = 80-100fL MCH = 27-32pg MCHC = 33-36% ▪ Shortens incubation time from 10-15min to 3min = dihydrogen potassium phosphate
40. Sources of Error in Automated Cell Counting: ▪ Non-ionic detergent = promotes RBC lysis
a. APERTURE PLUGS (most common), BUBBLES, EXTRANEOUS ELECTRICAL PULSES b. Cyanmethemoglobin is read at 540nm
causes POSITIVE error c. Cyanmethemoglobin method cannot detect: sulfhemoglobin
b. Hemolysis causes NEGATIVE ERROR d. Lipemia can be corrected using a patient blank.
c. Two cells suspended in the solution were counted as one, leading to increase in cell size 44. Hematopoiesis:
= COINCIDENCE a. Hematopoietic stem cell marker = CD34
d. Giant platelets = false decrease in platelet count b. Multi-colony stimulating factor = IL-3
e. Schistocytes (RBC fragments) = cause spurious decrease in RBC count c. Site of hematopoiesis in developing embryo = YOLK SAC → LIVER/SPLEEN → BONE
f. Platelet satellitism = false decrease in platelet count MARROW
g. RBC agglutination, cold agglutinins or clot = false decrease in RBC count d. Erythropoietin is produced by the KIDNEYS in response to TISSUE HYPOXIA
\ Quality Assurance and Control
41. e. In erythropoiesis, hemoglobinization becomes evident in RUBRICYTE stage
a. Bull’s eye in quality control chart indicates good accuracy and good precision. f. Maturation series of RBCs: Rubriblast → Prorubricyte → Rubricyte →
b. Trend occurs when the control value moves in the same direction (increases or Metarubricyte → Reticulocyte → Erythrocyte
decreases) for 6 consecutive days. g. Life span of RBCs = 120 days
c. Shift occurs when the control value is on one side of the mean for approximately 6 h. Pathway needed to generate ATP for the RBCs = EMBDEN MEYERHOF
consecutive days or is beyond 1SD on the same side of the mean for at least 4 i. Pathway needed to generate 2,3-DPG = RAPOPORT –LUEBERING
consecutive days. j. In leukopoiesis, primary (nonspecific/azurophilic) granules appear in =
d. Determination of the slope and Y-intercept is one of the most useful ways of comparing PROMYELOCYTE
methods. (p29 Barbara Brown) k. In leukopoiesis, Secondary granules appear in MYELOCYTE STAGE
▪ Slope represents proportional systematic error. If both procedures are in l. Host defense against parasitic infections; suppresses basophil reaction = EOSINOPHIL
complete agreement, the slope will be 1.0 m. Not an end cell: LYMPHOCYTES
▪ Y-intercept represents constant systematic error. Perfect correlation between n. Cell that exhibits a cartwheel like pattern on its nucleus, a large, well-defined hof and a
two procedures will give a Y-intercept value of 0 deeply basophilic cytoplasm = PLASMA CELL
42. Hemoglobin determination (Acid Hematin) 45. Microcytic, Hypochromic Anemia: Thalassemia, Iron deficiency anemia, Chronic blood loss,
a. Uses Sahli pipet (volume capacity of 20uL or 0.02mL) Sideroblastic anemia
b. Reagent: hydrochloric acid \ Sideroblastic anemias are a group of disorders characterized by defective iron loading
46.
subsequently leading to accumulation of iron in the mitochondria or erythroid precursors.
47. Thalassemia = decreased globin synthesis; quantitative globin defect

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48. Alpha thalassemia is characterized by: 60. FAB Classification of Acute Myeloproliferative disorders
a. HbA, HbA2 and HbF are DECREASED; a. M0 – acute undifferentiated leukemia
b. HbH (4-beta) and Bart’s Hgb (4-gamma) are INCREASED b. M1 – acute myeloblastic leukemia without maturation
49. Beta thalassemia (Cooley’s anemia) is characterized by: c. M2 – acute myeloblastic leukemia with maturation
a. HbA is DECREASED; HbA2 and HbF are INCREASED d. M3 – acute promyelocytic leukemia
50. Iron deficiency anemia = decreased serum iron, TIBC is INCREASED M3 is associated with Disseminated Intravascular Coagulation (DIC) and chromosomal
- .
51. Porphyria is a group of rare disorders caused by specific enzyme deficiency necessary for the abnormality involving t(15;17)

§
synthesis of heme. This leads to accumulation of porphyrins and/or their precursors. e. M4 – acute myelomonocytic leukemia
52. Megaloblastic anemia is a macrocytic, normochromic type of anemia. It is associated with f. M5 – acute monocytic leukemia (Schilling’s leukemia)
impaired DNA synthesis. Causes of megaloblastic anemia include: pernicious anemia, sprue, g. M6 – acute erythroleukemia (DiGuglielmo)
following gastrectomy, dietary, abnormal intrinsic factors, tapeworm infection (D. latum) h. M7 – acute megakaryocytic leukemia
53. Anemia of Endocrine disorders include anemia that developed secondary to interference 61. FAB Classification of Chronic Myeloproliferative disorders
with normal action of hormones on RBC production (erythropoiesis). Anemia may appear as a. Chronic Myelogenous Leukemia
a result of hypothyroidism, hypopituitarism, adrenal abnormalities and hypogonadism. ▪ 90-95% of patients with CML are positive for Philadelphia chromosomes; it
(Clinical Hematology: Principles, Procedures and Correlations by Steininger) indicates GOOD PROGNOSIS
54. Normocytic, Normochromic anemia includes: ▪ 5-10% of patients with CML are negative for Philadelphia chromosome; it indicates
a. Sickle cell anemia, Aplastic anemia, Acute blood loss POOR PROGNOSIS
b. Anemia associated w/chronic disorders, renal & endocrine disease b. Polycythemia vera
c. Metastatic cancer, Leukemia, Multiple myeloma ▪ Characterized by panmyelosis (increased RBC count, granulocyte count and platelet
d. Hodgkin’s disease; Leukoerythroblastosis count) and decreased erythropoietin levels
e. Paroxysmal nocturnal hemoglobinuria; HDN, Certain acquired hemolytic anemia c. Essential Thrombocythemia
55. Major cause of death in patients with sickle cell anemia = INFECTIOUS CRISES d. Myelofibrosis
56. Diamond Blackfan anemia is known as congenital pure red cell aplasia 62. FAB classification of Acute Lymphoproliferative disorders include:
-
57. Acute Leukemia a. L1 = small cell, homogenous; childhood
\ a. FAB = >30% blasts in peripheral blood b. L2 = large cell, heterogenous; adults
b. WHO = at least 20% blasts in peripheral blood c. L3 = Burkitt type
58. Chronic leukemia is characterized by <10% blasts in the peripheral blood 63. FAB classification of Chronic Lymphoproliferative disorders include:
59. Dymyelopoietic Disorders (Myelodysplastic or Pre-leukemia) include: a. Chronic lymphocytic leukemia (CLL)
a. Refractory Anemia (RA)/ Refractory Cytopenia (RC) b. Prolymphocytic leukemia
b. RA with ringed sideroblasts c. Hairy cell leukemia
c. RA with excess blasts
d. Chronic myelomonocytic leukemia
e. RA with blasts in transformation

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HEMOSTASIS 9. Methods for bleeding time = Duke’s method and Ivy method
10. Screening test for secondary hemostasis = CLOTTING TIME
1. Platelets are cells that bud off from the megakaryocytes. They have no nucleus and they have 11. Activates the extrinsic pathway = TISSUE FACTOR or TISSUE THROMBOPLASTIN
life span of 8-11 days. 12. Disseminated Intravascular coagulation
2. Maturation Sequence of platelets a. Increased PT, APTT, TT
a. Reddish blue granules, no budding thrombocytes = megakaryocytes b. Decreased factor I (within 4-24 hours) and decreased platelet count (up to 48 hours)
b. Aggregates of granular material, with budding thrombocytes (platelet shedding) is c. Positive D-dimer (within 4 hours)
visible = metamegakaryocytes 13. Manual Fibrin Clot Determination: WIRE LOOP METHOD AND TUBE TILTING
3. Platelet functions: Adhesion → Activation → Aggregation 14. Fibrometer is based on the principle of: ELECTROMECHANICAL METHODS
a. Requirements for Platelet adhesion: vWF and gpIb 15. Factors that deteriorate at room temperature: V and VIII
b. Requirements for Platelet aggregation fibrinogen and gp IIb-IIIa 16. Factors that are activated at refrigerated temperature (4°C) VII and XI
4. Platelet Structure 17. Adsorbed by barium sulfate = Prothrombin group (Vitamin K dependent factor).
a. Outer layer of the platelets = GLYCOCALYX \ Vitamin K dependent factors include factors II, VII, IX and X. First factor to be affected in
18.
b. Glycocalyx, plasma membrane and submembranous area belong to the PERIPHERAL vitamin K deficiency is factor VII.

:c.
d.
ZONE.
Microfilaments and microtubules are found in the SOL-GEL ZONE
Responsible for clot retraction = THROMBOSTHENIN (Actomyosin)
Protein S and Protein C are also vitamin K dependent.
19. In the Intrinsic pathway, IXa + Ca2+ + PF3 + VIIIa activates factor X
20. In the Extrinsic pathway, III + Ca2+ + VIIa activates factor X
e. Alpha granules, dense bodies and mitochondria = ORGANELLE ZONE 21. Prothrombin-converting factor (prothrombinase complex)
f. Dense bodies of platelet include: = Xa + V + Ca2+ + PF3
▪ ADP, ATP, GDP, GTP 22. Heparin is considered as an in vivo, in vitro anticoagulant.
▪ Calcium, Magnesium 23. Detects factor XIII deficiency = DUCKERT’S TEST (5M urea solubility test)
▪ Serotonin, Pyrophosphates 24. Reagents of prothrombin time = THROMBOPLASTIN, CALCIUM CHLORIDE
5. Platelet factor 3 = phospholipid 25. International Normalized ratio is computed in PT. Most responsive thromboplastin reagents
6. Coagulation factors that are not numbered: HMWK and PK have International sensitivity index near 1
7. Normal levels of fibrinogen range between 200-500mg/dL. If fibrinogen levels are extremely 26. Reagents of activated partial thromboplastin time = ACTIVATOR, PHOSPHOLIPID, CALCIUM
low (<100mg/dL), PT and aPTT may also be prolonged. CHLORIDE
8. Normal Value for Bleeding time = 2-8 minutes 27. Used to reverse heparin overdose = PROTAMINE SULFATE

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28. Tests that detect for deficiencies of the common pathway = STYPVEN TIME
29. Test for fibrinogen deficiency, not affected by heparin therapy = REPTILASE TIME
30. Stable factor is known as factor VII while labile factor is known as factor V.
31. Labile clotting factors include: factor V and factor VIII
32. Hemophilia C is factor XI deficiency.
33. Petechiae – purplish-red, pinpoint hemorrhagic spots (3mm in diameter) in the skin caused
by loss of capillary ability to withstand normal blood pressure and trauma
34. Purpura – purple skin discoloration of about 1cm in diameter seen in mucocutaneous
bleeding
35. Ecchymosis – are 3cm or large and usually irregular in shape; subcutaneous form of purpura
in which blood escapes into large areas of skin and mucous membranes, but not into deep
tissues; bruise
36. Principal enzyme involved in fibrinolysis = PLASMIN
37. Primary inhibitor to fibrinolysis = alpha-2-antiplasmin
38. Euglobulin clot lysis time is used to screen for = FIBRINOLYSIS
39. Enzyme inhibited by aspirin = CYCLOOXYGENASE
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FINAL COACHING
IMMUNOLOGY, SEROLOGY & BLOOD BANKING
1. History and Related Personalities:
a. Edward Jenner = demonstrated cross immunity by establishing relationship between
exposure to cowpox and immunity to small pox
b. Columbian theory suggests that syphilis was endemic in Haiti and was subsequently
contracted and carried to Europe by the crew of Christopher Columbus
c. ELIE METCHNIKOFF demonstrated phagocytosis and cellular immunity
d. Year of T cell receptor gene discovery = 1984
e. Clonal Selection theory = Frank Macfarlane Burnet
f. Radioimmunoassay was developed by Rosalyn Yallow. Gamma counters (solid
scintillation counters) are used to measure radioisotope labels during
radioimmunoassays.
g. Louis Pasteur developed live attenuated virus vaccine for rabies.
h. The Hybridoma technology is used to produce monoclonal antibodies. The significance of
this technique is reflected on the fact that a Nobel prize was awarded in 1984 to Georges
Kohler and Cesar Milstein for their work in this field.
i. Paul Ehrlich - antibody formation theory
j. Antibody Diversity = Susumu Tonegawa
k. Gerald Edelman and Rodney Porter for the structure of antibodies
l. George Snell, Jean Dausset and Baruj Benaceraf for the MHC (Major Histocompatibility
Complex)
m. Cook who spread a febrile disease (Salmonella) that killed almost 50 persons = Mary
Mallon. Carriers of Salmonella typhi can be tested using their stool specimen.
n. First recorded blood transfusion in history = Pope Innocent from 3 donors in the year
1492
o. 1901, Karl Landsteiner discovered the ABO blood group system. He wrote the book “The
Specificity of Serological Reactions”
p. 1943 = Loutit and Mollison introduced the use of acid-citrate-dextrose (ACD) as
preservative. Citrate in blood bags function as anticoagulant it acts by binding calcium.
q. 1957 = Gibson introduced citrate-phosphate-dextrose
r. Dr. Yves Lapierre developed the gel technology/test.
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2. Insufflation, variolation = the process of inhaling/injecting powder made from small pox 18. Interleukins
scabs to induce immunity/protection a. IL-1 = responsible for fever
3. Salk vaccine = formalin-killed poliovirus; injected, stimulates antibody production in serum b. IL-2 = promotes differentiation of T cells
but not in mucosa c. IL-3 = multi-colony stimulating factor; supports the growth of pluripotential stem cells
4. Sabin vaccine = live-attenuated poliovirus; orally administered; stimulates production of of the hematopoietic system
both IgA and IgG d. IL-8 = activating and chemotactic factor for neutrophils
5. Process by which cells are capable of moving from the circulating blood to the tissues by 19. Haptens are antigens that are not immunogenic on their own but will only be immunogenic
squeezing through the wall of a blood vessel = DIAPEDESIS when coupled to a carrier.
6. Monocyte-Macrophage System: 20. Proteins are most immunogenic. Lipids and nucleic acids are weakly immunogenic.
▪ in liver = Kupffer cells 21. Antigen-binding site is the PARATOPE. Antigen determinant is the EPITOPE.
▪ in kidneys = Mesangial cells 22. Hinge region is found in between the CH1 and CH2 region. It has high proline content which
▪ in CNS = Microglial cells allows for flexibility of the immunoglobulin. CH means constant heavy chain.
7. Functional groups of lymphocytes: T cells, B cells and Natural Killer cells 23. Differences in the constant region of the heavy chain = ISOTYPE
8. % of B cells in the blood: 20-30%. Most of the stimulated B cells are induced to become
plasma cells and some into memory B cells.
\
24. Antibodies/Immunoglobulins:
▪ IgM in plasma is pentameric and can bind to 10 antigens simultaneously.
9. Stimulates transformation of B cells into plasma cells = T HELPER CELLS ▪ Primary response antibody = IgM
10. Natural killer cells (null cells, large granular lymphocytes, 3rd population of lymphocytes) ▪ Secondary/Anamnestic response antibody = IgG
are part of the natural/innate immunity. ▪ Immunoglobulin that can cross the placenta = IgG
11. Perforins are found in natural killer cells and cytotoxic T cells. ▪ Immunoglobulin that is best in agglutination and complement activation = IgM
12. T cells and B cells are part of the adaptive/acquired immunity. ▪ Immunoglobulins found in the surface of B cell = IgM and IgD
13. Most effective antigen presenting cell = Dendritic cells ▪ Immunoglobulins that have J-chain = IgM and IgA2
14. Steps in phagocytosis: ICED (initiation, chemotaxis, engulfment, digestion/degradation) ▪ Immunoglobulins that can activate complement = IgM and IgG
15. Forms of Phagocytosis ▪ The secretory component of IgA2 protects the immunoglobulin from being degraded by
a. Direct = through PPRR (primitive pattern recognition receptors) proteolytic enzymes in mucosa/secretions.
b. Indirect = through opsonization 25. Pepsin split the immunoglobulin into 2 fragments. Papain splits the immunoglobulin
16. C3b, CRP and antibodies (immunoglobulin) act as opsonin. They coat the pathogen to molecule into 3 fragments.
enhance phagocytosis. 26. Svedberg unit (S) is used in expressing sedimentation coefficients in ultracentrifugation.
17. Chronic granulomatous disease is characterized by defective respiratory/oxidative burst. 27. Anergy is a term in immunobiology that describes a lack of reaction by the body's defense
Screening test for CGD include the neutrophil reduction test (aka Nitroblue tetrazolium dye mechanisms to foreign substances and consists of a direct induction of peripheral
test). lymphocyte tolerance. An individual in a state of anergy often indicates that the immune
system is unable to mount a normal immune response against a specific antigen, usually a
self-antigen.
28. Most common immunodeficiency disorder = SELECTIVE IgA DEFICIENCY

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29. Complement deficiencies
a. Most common complement deficiency = C2 deficiency
b. Most severe complement deficiency = C3 deficiency
c. C1. C2. C4 = Lupus-like syndrome
d. C3 = severe recurrent infections, glomerulonephritis
e. C5-C8 = Neisseria infections
f. C9 = NO KNOWN DISEASE ASSOCIATION
g. C1 INH = hereditary angioneurotic edema
30. Deficiency of MIRL and/or DAF can be associated with Paroxysmal nocturnal
hemoglobinuria (PNH). MIRL is membrane inhibitor of reactive lysis (also known as CD59)
while DAF is decay accelerating factor (also known as CD55).
31. Classical pathway of complement is activated by the presence of antigen-antibody complex
-
40. One of the most frequently used methods for obtaining lymphocytes is density gradient
centrifugation with Ficoll-Hypaque. Diluted defibrinated or heparinized blood is carefully
layered on top of the solution, and the tube is centrifuged. Centrifugation produces three
distinct layers: plasma at the top of the tube, a mononuclear layer banding on top of the
density gradient solution, and erythrocytes and granulocytes at the bottom of the tube
41. HLA Class I molecules present antigens to CD8+ cells (cytotoxic T cells). HLA Class II
molecules present antigens to CD4+ cells (helper T cells).
42. Mixed leukocyte culture or Mixed Lymphocyte Reaction is a cellular approach to tissue
matching that mimics in vivo condition of transplantation. It is based on the response of one
cell in tissue culture to the alloantigens on the surface of a second cell. It is considered an in
vitro measure of class II disparity between individuals that recognizes determinants found
on class II molecules. It is performed to determine whether the mononuclear cells of a

32. Calcium stabilizes the C1qrs complex during the recognition phase of the classical pathway prospective tissue transplant recipient will react against a potential donor’s leukocyte
of complement cascade. antigens. Heparinized blood samples are usually collected from both the prospective donor
33. Fragments a and b in complement cascade indicates that the complement component has and recipient.
been fragmented (cleavage). 43. Principle of microlymphocytotoxicity by dye exclusion: Cells and serum are incubated
34. C3 convertase: together in the wells of a microtiter tray. Mineral oil at the top of the wells prevents fluid
a. Classical pathway = C4b2a evaporation. Complement is added, and a second incubation is performed. The cells are
b. Alternative pathway = C3bBb washed, eosin is added, and the cells are fixed with formalin. Cells that fixed antibody and
35. Properdin stabilizes the C3bBb complex. were killed through the action of complement take up eosin and appear dark and
36. Membrane Attack Complex is made up of: C5b6789 nonrefractive under a phase-contrast microscope. In contrast, living cells are refractive and
37. Complement fragments can be measured via RADIAL IMMUNODIFFUSION. are easily identified by phase contrast microscopy. Variations of the
38. CD Markers: microlymphocytotoxicity assay are used for human leukocyte antigen (HLA) typing, HLA
a. CD2 = sheep RBC receptor crossmatch, and detection of anti-HLA antibodies.
b. CD4 = helper T cell, receptor for HIV 44. HLA and disease associations
c. CD8 = cytotoxic T cell a. Ankylosing spondylitis (or Bamboo spine disease) = HLA-B27
d. CD21 = receptor for EBV b. Rheumatoid Arthritis = HLA-DR4
39. The rosette technique using red blood cells from sheep has been a historical method for c. Systemic Lupus Erythematosus = HLA-DR3, DR2
enumeration of T lymphocytes. Lymphocytes are separated from whole blood and then mixed d. Graves Disease = HLA-B8 and DR3

i
with a suspension of sheep red blood cells. If three or more red blood cells are attached to a 45. Examples of type I (immediate/allergic) hypersensitivity reactions include hay fever,
lymphocyte, it is considered a rosette. Sheep cells attach to the CD2 antigen (sheep RBC asthma and food allergies.
receptor), found only on T cells. Using a counting chamber, 200 cells are counted and the 46. Examples of type III (immune complex) hypersensitivity reactions include SLE, rheumatoid
-
percent forming rosettes is calculated. This represents the percentage of T cells. arthritis, Arthus reaction and Serum sickness.

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47. Examples of type IV (delayed) hypersensitivity reaction include\ tuberculin test andIcontact
dermatitis.
48. Autoantibodies and associated autoimmune diseases:
a. Anti-smooth muscle antibodies = chronic active hepatitis.
b. Anti-mitochondrial antibodies = primary biliary cirrhosis
c. Anti-microsomal antibodies = Hashimoto’s thyroiditis
d. Anti-TSH receptor antibodies = Graves disease
49. Autoantibodies present in SLE patients include anti-double stranded DNA (ds-DNA)
antibodies, anti-nuclear antibodies (ANA), anti-extractable nuclear antigens (ENA), anti-
leukocyte antibodies, anti-Smith antibodies and anti-phospholipid antibodies (also known
as Lupus anticoagulant, which can cross the placenta and cause spontaneous abortion).
50. Fluorescein isothiocyanate (FITC) emits green fluorescence and phycoerythrin (PE) shines
red after excitation.
51. One particularly sensitive assay for ds-DNA is an immunofluorescent test using Crithidia
luciliae, a hemoflagellate, as the substrate. This trypanosome has circular ds-DNA in the
kinetoplast
52. A speckled pattern in ANA (anti-nuclear antibody) test can sometimes be found in
rheumatoid arthritis or lupus. Therefore, if a speckled pattern is obtained, more specific
testing such as an immunodiffusion assay should be done. Presence of anti-Smith antibody
would be diagnostic for lupus.
53. Sjögren's syndrome (SS) is a chronic autoimmune inflammatory disorder characterized by
diminished lacrimal and salivary gland function leading to dryness of eyes and mouth
(decreased tears and saliva).
54. Rheumatoid factor is an IgM autoantibody against the Fc portion of IgG.
55. In Rheumatoid Factor Latex tests:
a. Latex coated with human immunoglobulin (IgG) is used as reagent
b. a titer of 80 or greater is generally considered a positive result. A titer of 20 to 40 is
considered weakly positive.
c. No agglutination at 1:20, the specimen should be considered negative (even if
subsequent dilution shows agglutination)
56. Anti-human CRP antibodies attached to the latex in the CRP latex agglutination tests are
derived from GOAT OR RABBITS
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57. Steps in pregnancy testing using agglutination inhibition method:
a. Step #1 Addition of anti-hCG antibodies
b. Step#2 Addition of hCG-coated latex particles
s
c. Positive result: absence of agglutination
58. A false-negative reaction may take place in the prozone due to high antibody concentration.
If it is suspected that the reaction is a false negative, diluting out antibody and performing
the test again may produce a positive result.
59. In the postzone, excess antigen may obscure the presence of a small amount of antibody.
Typically, such a test is repeated with an additional patient specimen taken about a week
later. This would give time for the further production of antibody.
60. The reciprocal of the last dilution still exhibiting a visible reaction is the titer, indicating the
antibody’s strength.
61. ASO Titration/Tube test
a. The two main damaging sequelae to group A streptococcal infections are acute
rheumatic fever and poststreptococcal glomerulonephritis
b. ASO titration/tube test is based on neutralization/inhibition principle.
c. ASO Titer is reported as the reciprocal of the highest dilution demonstrating no
hemolysis
d. Normal: <200 IU
e. SLO control should show COMPLETE HEMOLYSIS
f. RBC control should show NO HEMOLYSIS
62. Heterophil Antibody Tests
a. Davidsohn Differential Test is a classic/traditional differential test to distinguish
between heterophil sheep cell agglutinins in human serum due to Forssman antigen,
serum sickness and infectious mononucleosis. Sheep RBCs are used in the agglutination
procedure, after adsorption with guinea pig kidney cells and beef RBCs.
▪ Forssman Antibodies– adsorbed by GPK cells only
▪ Infectious Mononucleosis– adsorbed by Beef or ox RBCs only
▪ Serum Sickness– adsorbed by GPK cells and Beef RBCs
b. Paul-Bunnell Test is a traditional test for heterophil antibodies, uses SHEEP RBCs
c. Monospot is a rapid test for heterophil antibodies; uses HORSE RBCs
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63. Slide agglutination for Salmonella (Widal agglutination test) 70. The yeast Saccharomyces cerevisiae is used to produce hepatitis B vaccine. The gene (a
a. A single rapid slid test is not considered positive unless a value of at least “+2” is segment of DNA strand) which controls the production of small HBV fragments is first
assigned in a sample taken in the first 2-3 weeks of infection. inserted into the microorganism. The yeast then produces virus fragments which are
b. Grading: subsequently collected, purified and used as a vaccine.
▪ 0 = no agglutination 71. Immunofixation electrophoresis is used to identify Bence Jones protein in patients with
▪ +1 = 25% agglutination multiple myeloma
▪ +2 = 50% agglutination 72. Bence-Jones proteins are abnormal, identical, monoclonal light chains that are present in
▪ +3 = 75% agglutination the urine of patients with multiple myeloma. They precipitate at 40-60°C but dissolves
▪ +4 = 100% agglutination when temperature reaches 100°C.
64. Coagglutination uses bacteria as inert particles to which antibody is attached. 73. Inhibition Immunofluorescent assay is a blocking test in which antigen is first exposed to
Staphylococcus aureus is most commonly used due to the presence of protein A on its unlabeled antibody, followed by the labeled antibody. It is then washed and examined. If the
surface, which is capable of naturally absorbing the Fc portion of antibody molecules. This unlabeled and labeled antibodies are both homologous to the antigen, there should be no
has been used in identification of Streptococci, Neisseria. Vibrio cholera 0139 and fluorescence.
Haemophilus influenzae. 74. Southern blot involves the analysis of DNA fragments using probes. This is also known as
the Southern Hybridization Asssay. Forthem blot RNA ; Western blot gpsp ; Eastern blot l
-

91%941
""
65. Hemagglutination inhibition – similar to agglutination inhibition, except that antigen-coated -
-

-
.
.
-
-

RBCs are added instead of antigen-coated latex particles; used for detection of HBs antigen
in hepatitis
66. Viral hemagglutination – used in viruses that can be quantified based on their ability to
agglutinate red blood cells, such as influenza virus
67. Radial immunodiffusion is an example of single immunodiffusion technique; whereas
Ouchterlony method is an example of a double immunodiffusion technique.
68. ONCOFETAL ANTIGENS are markers most highly expressed in both normally developing
75. Quantitative tests are performed by RT-PCR, real-time PCR, or branched DNA amplification
in order to monitor the amount of HCV RNA or viral load by patients before, during or after
antiviral therapy in chronically infected individuals.
76. Polymerase chain reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP)
are examples of molecular assays.
77. Polymerase Chain Reaction is a DNA target amplification technique. It uses heat stable Taq
-
polymerase, primers to copy the target DNA.
-

fetal tissue and in certain kinds of cancers. Examples of oncofetal antigen include CEA and 78. Steps in Polymerase chain reaction: Denaturation → Annealing →
-
AFP. Extension/Elongation/Synthesis
-
69. Tumor Markers:
\ 79. The gold standard for the diagnosis of leptospirosis is the microscopic agglutination test
y
a. CYFRA 21-1 = lung C. (MAT).
Ib. BTA and NMP = bladder C. 80. Rapid Enzyme immunoassay (EIA) test for Helicobacter pylori (which causes peptic ulcers)
y
\c. CA 27-29 = breast C. detects for antigen in stool specimen.
d. BRCA-1, BRCA-2, HER2/neu = breast C. 81. NS1 antigen is used for evaluation of Dengue virus infection.
e. CA 125 = ovary C. 82. OptiMal assay detects parasitic lactate dehydrogenase produced by viable malaria parasite
f. CA 19.9 = pancreatic/gastric C. ; assoc with Lewis 83. MalaQuick Standby Malaria test detects for HRP-2 (histidine-rich protein-2) produced by
.

system
g. Calcitonin = medullary thyroid C.
\ Plasmodium falciparum.
\h. Beta-HcG = testicular C.

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84. Immunogenic Entamoeba histolytica antigens 91. Reagents in VDRL

a. Serine-rich Entamoeba histolytica protein (SREHP) a. 0.03% cardiolipin = phospholipid isolated from beef heart; responsible for reactivity
b. Lectin protein (adhesion) b. 0.9% cholesterol = serves as center for absorption of tissue lipids to increase Ag size
c. Cysteine-rich protein c. 0.21% lecithin = to produce standard reactivity
85. Frei test is a skin test for lymphogranuloma venerum, which is caused by Chlamydia 92. Complement inactivation is done at 56degC for 30 mins. When more than 4 hours have
trachomatis. elapsed from the inactivation, a reinactivation is performed by heating the serum at 56°C
86. Casoni Skin test is an intradermal test for Echinococcus granulosus, the causative agent of for 10 minutes.
hydatid disease 93. Advantages of Rapid Plasma Reagin
87. Schick test is a serological susceptibility test used to determine the presence of immunity to a. No requirement for heat inactivation
Corynebacterium diphtheriae, which is the causative agent of diphtheria. It was named after b. Antigen suspension more stable, can be stored
Bela Schick, a Hungarian-American pediatrician. c. Charcoal makes reaction visible, thus results are read macroscopically
88. Dick test is a serologic test used to identify those susceptible to scarlet fever (caused by d. Test can be automated, more sensitive and rapid than VDRL
Streptococcus pyogenes). 94. Treponemal Tests for Syphilis
89. Non-treponemal tests for Syphilis a. Detects for: anti-Treponema pallidum antibodies
a. Detects for: anti-cardiolipin antibodies (aka. Antibodies to Wasserman antigen, or b. Main Reagent: Treponema pallidum
reagin). c. Examples:
b. Main Reagent: Cardioplipin ▪ TPI (Treponema pallidum Immobilization)
c. Examples: ▪ FTA-Abs (Fluorescent Treponemal Antibody-Absorption)
▪ Unheated Serum Reagin (USR) ▪ Agglutination Assays (TPHA, MHA-TP, HATTS, TPPA)
▪ Toluidine Red Unheated Serum Test (TRUST) 95. Interpretation: Treponema pallidum Immobilization
▪ Reagin Screen Test and Automated Reagin Test a. Positive: >50% immobilized
▪ Wasserman Complement Fixation Test b. Negative: <20% immobilized
▪ Kahn Flocculation Test, c. Doubtful: 20-50% immobilized
▪ Plasmacrit 96. Treponemal Agglutination Tests
▪ RPR (Rapid Plasma Reagin) a. TPPA (Treponema pallidum particle agglutination)
▪ VDRL (Venereal Disease Research Laboratory) test - Gelatin particles coated with treponemal antigens
90. Non-treponemal tests – Rotators b. TPHA (Treponema pallidum hemagglutination)
a. VDRL = 180 RPM for 4 mins - tanned sheep red cells coated with antigen from the Nichols strain
b. RPR = 100 RPM for 8 mins c. MHA-TP (Microhemagglutination-Treponema pallidum)
- Microtechnique; formalinized, tanned sheep erythrocytes sensitized with
Treponema pallidum antigen (Nichols strain)
d. HATTS (hemagglutination treponemal test for syphilis)
- glutaraldehyde-stabilized turkey RBCs

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97. One of the most used confirmatory tests is the FTA-ABS test, an indirect fluorescent
antibody test. A dilution of heat-inactivated patient serum is incubated with a sorbent
consisting of an extract of nonpathogenic treponemes (Reiter strain). This removes cross-
reactivity with treponemes other than T. pallidum. It uses dead Treponema pallidum in
slide (Nichols strain) as antigen.
98. BLOOD is considered both as a biologic and as a DRUG.
99. ABO blood group is ISBT 001. Rh blood group is ISBT 004.
100. Anti-A reagent has BLUE color. Anti-B reagent has YELLOW color due to the presence of
acroflavin dye. Anti-human globulin (AHG) reagents are often GREEN in color.
101. Group I discrepancy = weak or missing antibodies (newborn and elderly)
102. Individuals with Acquired B antigen will appear to be blood type AB during the forward
typing but will appear as blood type A in the reverse type. It belongs to group II
discrepancy.
103. An A2 subgroup will react with anti-A, leading to a positive reaction but will produce a
negative reaction with anti-A1 lectin (Dolichos biflorus).
Blood Reaction with Reaction with anti-A1
Group A Anti-A reagent lectin (Dolichos biflorus)
A1 Positive Positive
A2 Positive Negative
104. PUBLIC ANTIGEN to an antigen characteristic of the RBC membrane found commonly
among individuals, often in more than 98% of the population. PRIVATE ANTIGEN is an
antigen characteristic of the RBC membrane that is unique to an individual or a related
family of individuals, and is therefore not commonly found on all cells. They often exist only
in less than 1% of the population
\
105. Mendel’s 1st Law of Inheritance: Law of Segregation
106. Homozygous means that an individual has two of the same allele. Heterozygous means that
an individual has one each of two different allele.
107. A heterozygous A mother and heterozygous B father can have children of blood types A, B,
AB and O.
108. Rh-positive mom and Rh-positive dad can have Rh negative baby if they are both
heterozygous for the D (Dd).
109. Rh-negative mom and Rh-negative dad can only have Rh-negative offspring.
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110. Type 1 chain precursor is formed when galactose on the ABH precursor substance is
attached to the N-acetylglucosamine in the beta-1-3 linkage. Type 2 chain precursor is
formed when galactose of the ABH precursor is attached to the N-acetylglucosamine in the
beta 1-4 linkage.
111. Elution refers to removal of antibodies bound to the antigens on RBCs. Cells that are coated
with antibody are treated in such a manner as to disrupt the bonds between the antigen and
antibody. This antibody serum then can be tested to identify its specificity using routine
methods. The mechanism to free the antibody may be physical (heating, shaking) or
chemical (ether, acid). The harvested antibody-containing fluid is called eluate.
112. Fluids in which A, B and H substances can be detected in secretors include: saliva, tears,
urine, digestive juices, bile, milk, amniotic fluid and pathologic fluids such as pleural,
peritoneal, pericardial and ovarian cyst fluids.
113. Lewis antigens are manufactured by tissue cells, secreted into the body fluids and is
adsorbed onto the RBCs from the plasma. They are neutralized by secretions such as saliva.
Lewis antibodies are mostly IgM and are unlikely to cause HDN.
114. One of the mechanisms that cause weak D phenotype includes the D mosaic or the partial D
formation. This causes a patient to be typed as D-positive and at the same time, capable of
producing an allo-anti-D which has the ability to react with D-positive red blood cells other
than their own. Therefore, recipients that are of the D mosaic phenotype should be
considered as_RH NEG (D NEG), and be given Rh-negative blood to prevent transfusion
reaction to prevent the reaction of allo-anti-D from the recipient with the D-antigen from
the donor.
115. Most reactions produce optimal antigen–antibody combination when the pH is between
6.5-7.5, but there are some exceptions, such as human anti-M, which react best at a lower
pH.
I
116. Cord cells are washed 6-8x (or 5x) to remove Wharton’s jelly, which is a viscous,
-
mucopolysaccharide material present on cord blood cells that can interfere with the
analysis.
117. Washing of RBCs in an AHG procedure removes the free/unbound antibodies which might
neutralize the AHG reagent. Failure to wash RBCs in an AHG procedure can lead to FALSE
NEGATIVE results. Washing must be done at least three times.
118. AHG can either be monospecific (contains antibodies to IgG or C3d) or polyspecific
(contains antibodies to both IgG and C3d)
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119. Polyclonal antibodies used in immunohistochemical stains are derived from rabbits.
120. Direct Antiglobulin Test detects in vivo sensitization. EDTA-anticoagulated blood samples
are preferred. DAT is positive in cases of hemolytic transfusion reaction, hemolytic disease
of the newborn, autoimmune hemolytic anemia
121. Pokeweed mitogen (PWM) is a mitogen that can polyclonally activate both the B cell and the
T cell.
122. Blood Goup O has the greatest amount of H antigen
\
123. Bombay individuals:
a. Serum of Bombay individuals contain anti-A, anti-B, anti-AB and anti-H. Serum
agglutinates with A cells, B cells, AB cells and O cells.
b. RBCs of Bombay individuals do not have H antigen. Bombay RBCs do not
agglutinate/react with anti-H (Ulex europaeus) lectin
124. Parabombay individuals possesses hh gene and Se gene. They are similar to Bombay
phenotype except that there is normal ABH expression in secretions and body fluids.
125. McLeod phenotype is very rare. All who have it are male, and inheritance is X-linked
through a carrier mother. McLeod phenotype RBCs lack Kx and Km and have marked
depression of all other Kell antigens. A significant proportion of the RBCs in individuals with
McLeod phenotype are acanthocytic. It has also been linked to CGD (chronic granulomatous
disease)
126. Patients with Duffy null phenotype forms anti-Fy3. Those with Jk null phenotype forms anti-
Jk3.
127. Jk null phenotype is abundant among Polynesians. It has also been found in Filipinos,
Indonesians. Chinese and Japanese.
128. Kidd antibodies have a notorious reputation in the blood bank. They demonstrate dosage,
are often very weak and are found in combination with antibodies, all of which make them
difficult to detect. Titer of Kidd antibodies quickly declines in vivo is the major reason why
they are a common cause of delayed hemolytic transfusion reaction.
129. The name Lutheran blood group was derived from the donor Luteran from whom the
antibody was first identified.
130. Auberger antigens belong to the Lutheran blood group system.
131. Indian blood group system was named as such because the first Indian positive individual
was first seen in India. Highest percentage of Indian positivity is found in Arabs.
132. Anti-U is formed in S-s- individuals.
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133. DIEGO blood group system is associated with Southeast Asian ovalocytosis and serves as an
anthropologic marker of Mongolian ancestry
134. Length of incubation with LISS is 10-15 mins. LISS acts by decreasing the zeta potential;
whereas polyethylene glycol acts by removing water.
135. Length of centrifugation (serofuge), during immediate spin phase is 20sec
136. Apheresis is derived from Greek word “aphairos” which means “to separate or remove or to
take from”
137. During erythrocytapheresis, two standard units of RBCs can be collected, or one unit of
RBCs collected concurrently with plasma and/or platelets.
138. Donors for plateletpheresis should have a platelet count of at least 150 x 103/uL. Donors for
platelet pheresis can donate after 2 days (48 hours) interval.
139. Resulting signs and symptoms of acute citrate toxicity can include perioral paresthesias
(tingling or numbness of lips/mouth), acral paresthesias, shivering, lightheadedness,
twitching, and tremors. As the ionized calcium levels fall farther, these symptoms may
progress to carpopedal spasm, tetany, and seizure
140. Acute normovolemic hemodilution is a type of autologous donation where the blood is
collected before a surgical procedure and simultaneously replaced with a comparable
volume of crystalloid or colloid solution. Blood collected during hemodilution can be saved
and retransfused as fresh whole blood.
141. During blood collection, the blood pressure cuff must be inflated to 40-60mmHg.
142.
\ Jet-like, pulsating, bright red blood during donation is a sign of arterial puncture during
blood donation. i immediately stop donation then apply
pressure
143. In copper sulfate method for assessing hemoglobin levels, an acceptable blood drop should
sink within 15 seconds in a copper solution with a specific gravity of 1.053.
- y
144. 2-week deferral for donors who have received a live attenuated or bacterial vaccine such as
measles (rubeola), mumps, polio, typhoid or yellow fever. 4-week deferral for donors who
have received a live attenuated vaccine for German measles (rubella) or chicken pox
145. No deferral for toxoids or killed or synthetic viral, bacterial or rickettsial vaccines if the
donor is symptom-free and afebrile.
146. During donor screening, both arms of the donor must be examined for signs of repeated
parenteral entry, especially multiple needle puncture marks. Such evidence is reason for
indefinite exclusion, since this may indicate IV drug use.
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147. Hydroxyethyl starch (HES) is used as an RBC sedimenting agent during leukapheresis in
s i -
order to separate WBCs from RBCs effectively.
148. Granulocyte concentrate should contain at least 1.0 x 1010 or 10 x 109 PMNs per unit.
149. Cryoprecipitate contains at least 80 IU of factor VIII:C and at least 150mg of fibrinogen/unit.
150. Cryosupernatant fluid is the FFP with reduced cryoprecipitate. Cryosupernatant contains a
variable amount of fibrinogen and a full complement of other coagulation factors apart from
factor VIII and factor XIII. Cryosupernatant is devoid of FVIII/vWF multimers.
151. Frozen RBCs are red cell products that can be stored for up to 10 years at -65 degrees
centigrade or colder temperature. It is indicated for patients with rare phenotypes, for
autologous donation as well as in national emergencies in which blood cannot be
dispositioned out of hospitals quickly enough to prevent expiration. They can be prepared
using cryoprotective agents such as hydroxylethyl starch or glycerol. High glycerol method
or the slow, controlled freezing process, uses 40%w/v glycerol and frozen using a mechanical
freezer which provides storage temperature of -80°C. Low glycerol method, which is known
as a rapid, more controlled freeze technique, uses 20% w/v glycerol and frozen using liquid
nitrogen (-120°C).
152. Irradiated blood inactivates T cells which eventually helps prevent the graft-vs-host-
disease.
153. Leukoreduction of an RBC unit requires the unit to have at least 85% RBC recovery rate.
Leukoreduced RBCs can be used to prevent febrile transfusion reaction, transfusion related
acute lung injury (TRALI) and CMV transmission.
154. A one-degree centigrade rise in the temperature of the patient from the baseline
temperature, occurring during transfusion is known as Febrile transfusion reaction.
155. Anaphylactic reactions are caused by anti-IgA antibodies. Washed RBCs are preferred to
recipients with history of anaphylactic reactions.
156. Hives and itching are signs of allergic reactions (Type I hypersensitivity)
157. Transfusion-associated circulatory overload (TACO) is a good example of an iatrogenic
(physician-caused) transfusion reaction. Patients at significant risk include children, elderly
patients, and patients with chronic normovolemic anemia, cardiac disease, or sickle cells.
\
158. Anticoagulants and corresponding shelf-life -
Open system transfuse Min 24 hrs
-
-
a. ACD, CPD, CP2D – 21 days
b. CPDA1 – 35 days
c. AS (Additive Solutions) – additional 7 days
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159. Packed RBC unit prepared using an open system must be transfused within 24 hours.
160. Pooled platelets must be transfused within 4 hours.
161. Any transfusion must be completed within 4 hours.
162. Febrile and cold agglutinins are antibodies that cause RBCs to aggregate at high or low
temperatures, respectively. They are believed to be a result of infection by organisms with
antigenic groups similar to some of those found on the RBCs.
163. Anti-I is related to Mycoplasma pneumoniae infection (Primary atypical
pneumonia/walking pneumonia). It is the most commonly encountered cold agglutinin.
164. Anti-i is related to Epstein-Barr virus infection, which causes infectious mononucleosis.
165. Majority of the IgG antibodies detected in eluates prepared from the WAIHA patient’s RBCs
or in the patient’s serum has a complex Rh-like specificity. Occasionally, an autoantibody
may have what is termed as “simple anti-e specificity”.
166. VS and f are antigens of the Rh blood group system. LW was originally under the Rh blood
group but is now a separate system, known as Landsteiner-Wiener system, ISBT 016.
167. Paroxysmal cold hemoglobinuria (PCH) is the least common type of autoimmune hemolytic
anemia (AIHA). In PCH, a biphasic IgG hemolysin causes red cell destruction. It reacts/binds
with red cells at cold temperature and lyses RBCs at warm temperature. The biphasic
hemolysin in PCH has anti-P specificity. It can be demonstrated in the laboratory through
the Donath-Landsteiner test.
\ Exchange transfusion is done in babies suffering from HDN. It can be used to remove excess
168.
bilirubin in baby’s blood.
169. Neocyte transfusion is indicated in transfusion-dependent patients, such as those with
sickle cell anemia and thalassemia.
170. ABO incompatibility causes the most severe hemolytic transfusion reaction. Rh HDN (anti-
D) causes the most severe hemolytic disease of the newborn (HDN).
171. Replacement for minor cross-matching = antibody screen
172. In emergency cases where the blood time of the patient/recipient could not be determined,
it would be best to transfuse O, Rh negative packed RBCs.
173. Grading agglutination Reactions (Harmening 5th)
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HISTOTECHNIQUES 10. Bouin‘s and Brasil‘s solutions are picric acid-containing fixatives.
11. Alcoholic iodine solution is used to remove excessive mercuric fixatives that may form when
using Zenker‘s solution.
1. Tissue processing follows the following order:
12. Microanatomical fixatives include: 10% formol saline, 10% neutral buffered formalin,
▪ Fixation
Heidenhain’s Susa, Formol sublimate, Zenker’s solution, Helly’s solution, Bouin’s solution
▪ Dehydration
and Brasil’s solution.
▪ Clearing (dealcoholization)
13. 10% neutral buffered formalin is a general-purpose fixative.
▪ Impregnation (infiltration)
14. The best fixative for cytologic smears is the ether alcohol (equal parts of 95%ethyl alcohol
▪ Embedding (blocking/ casting)
and ether). However, this has been abandoned because of the flammability, volatility and fire
▪ Section-cutting
hazards associated with ether. Most laboratories now use 95% ethanol for routine
▪ Staining
preparations that require fixation.
▪ Mounting
15. Acetic acid is normally used in conjunction with other fixatives to form a compound solution.
2. The first and most critical step in histotechnology involves fixing or preserving the fresh
It solidifies at 17 °C. It fixes and precipitates nucleoproteins, chromosomes and chromatin
tissue for examination. This process is known as fixation.
materials. It is very useful in the study of nuclear components of cells. It is an essential
3. Prosector is a person who dissects corpses for examination or anatomical demonstration.
constituent of most compound nuclear fixatives.
Diener refers to laboratory workers who assist in the performance of autopsies and
16. The rate of decalcification will depend upon the structure of tissues, temperature and
maintenance of morgues.
volume of the solution to be used. High concentrations and greater amount of fluid will
4. Brittle or hard tissues may be due to: prolonged fixation, prolonged dehydration, prolonged
increase the speed of the process
clearing, prolonged paraffin infiltration, overheated paraffin wax or drying out of tissues
17. The decalcifying fluid should be changed every 24-48 hours.
before actual fixation.
18. Tissues that should be decalcified include: bones, tuberculous organs and atherosclerotic
5. The maximum effectiveness of fixation is noted to be 20 times that of the tissue volume
vessels.
6. Fixation is enhanced by the use of smaller and thinner tissues, presence of agitation and
19. Carnoy’s fluid is considered as the most rapid fixative.
presence of moderate heat.
20. Phloroglucin-Nitric acid is the most rapid decalcifying agent.
7. An ideal fixative must be: cheap; stable; safe to handle; can kill cell quickly to prevent
21. Acid decalcifying agents include nitric acid, hydrochloric acid, formic acid, trichloroacetic
distortion; inhibit bacterial decomposition and autolysis; produce minimum shrinkage of
acid, sulfurous acid, chromic acid and citric acid.
cells; permit rapid and even penetration; harden tissues but not much; isotonic; able to make
22. Tissues that should be decalcified include: bones, tuberculous organs and atherosclerotic
cellular components insoluble to hypotonic solutions and render them insensitive to
vessels.
subsequent processing; and able to permit subsequent application of many staining
23. Determination of the completeness of decalcification include physical or mechanical
procedures.
methods, chemical test, or radiologic tests.
8. Methanol added as a preservative to formaldehyde will prevent its decomposition to formic
24. An ideal clearing agent: (1) should be miscible with alcohol; (2) should be miscible with
acid or precipitation to paraformaldehyde, but it serves to denature protein, thereby
paraffin wax; (3) should not produce excessive shrinkage, hardening or damage of tissue; (4)
rendering formalin unsuitable as a fixative for electron microscopy.
should not dissolve out aniline dyes; (5) should not evaporate quickly; and (6) should make
9. Most common metallic fixative: mercuric chloride
tissue transparent.

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25. Xylene turns milky when there is incomplete dehydration of the tissue.
26. Dioxane is a dehydrating and clearing agent.
27. Chloroform is the recommended clearing agent for tough tissues (e.g. skin, fibroid and
decalcified tissues), for nervous tissues, lymph nodes and embryos because it causes
minimum shrinkage and hardening of tissues
28. Paraffin is the simplest, most common and the best embedding medium used for routine
tissue processing
29. Common waxes have melting points of 45°C, 52°C, 56°C and 58°C. The 56°C wax is normally
used for routine work.
30. The paraffin oven should be maintained at a temperature 2 to 5°C above the melting point of
the paraffin wax to be used for impregnation.
31. When the sections needed have been cut from a paraffin block, it is recommended that it be
resealed to prevent drying of the tissue or destruction by insects. Resealing also makes
subsequent cutting easier.
32. Deparaffinization of tissues sections, especially prior to staining includes that passage of
tissues section to changes of xylene (xylol). Rehydration of tissues sections is accomplished
by subsequent exposure of deparaffinized tissues to decreasing alcohol concentrations prior
to washing with water and staining.
33. Most rapid embedding technique is the vacuum type.
34. Plastic embedding medium are classified into epoxy, polyester or acrylic based on their
chemical composition. Epoxy plastics used in microscopy includes Araldite, Epon and Spurr.
Polyester plastics are now seldom used. Acrylic plastics include polyglycol methacrylate and
methyl methacrylate.
35. Sectioning is process whereby tissues are cut into uniformly thin slices or sections with the
aid of a machine (microtome), to facilitate studies under the microscope
36. Clearance angle = 5-10°
37. The angle formed between the cutting edges, (bevel angle) is about 27-32°.
38. Sides of the wedge knife are inclined at an angle of about 15°.
39. Thickness of tissue sections
• Paraffin sections = 4 to 6μ
• Ultrathin sections = 0.5μ
• Celloidin sections = 10 to 15μ
• Disposable blades = 2 to 4μ
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40. Causes of an alternately thin and thick include: (1) blunt knife; (2) knife that is not clamped
properly; (3) when the tilt of the knife is too great; (4) when the knife or block holder is
loose; or when (5) knife tilt is too small that block is compressed by bevel and section is not
cut.
41. Sections are removed in ribbons of tens to allow easy location of serial sections. The sections
are then floated out on a waterbath set at 45-50 °C, approximately 6-10 °C lower than the
melting point of the paraffin wax used for embedding the tissue. This is to flatten the
sections and prepared them for mounting into the slides.
42. The purpose of stropping is to polish and sharpen the cutting edges while that of honing is to
remove the irregularities from the knife. A paddle strop made up of the best quality horse
leather, firmly attached to a solid back, in order to prevent sagging is preferred for use.
Honing Stropping
removal of gross nicks and removal of burrs or irregularities
blemishes on knife edge formed during honing
edge first edge last
heel to toe toe to heel
10 to 20 strokes on each side 40 to 120 double strokes
43. The cryostat is a refrigerated apparatus used in fresh tissue microtome, for freezing the
tissue into the block holder to the correct degree of hardness to facilitate easier and faster
sectioning. It consists of a microtome, usually a rotary microtome, kept inside a cold
chamber which is maintained at a temperature between -5 to -30 ºC (average of -20 ºC) by
an adjustable thermostat, capable of freezing fresh tissues within 2-3 minutes and cutting
sections of 4 µ with ease.
44. Common applications of frozen sections include: (1) rapid pathologic diagnosis during
surgery; (2) diagnostic and research enzyme histochemistry; (3) diagnostic and research
demonstration of soluble substances such as lipid and carbohydrates; (4)
immunofluorescent and immunocytochemical staining; (5) some specialized silver stains
45. Blueing is the procedure wherein decolorized sections are placed on an alkaline solution in
order to neutralize the acid and free the OH group, to form an insoluble blue aluminum
hematin-tissue lake. Blueing agents include warm tap water (40-50 °C), lithium carbonate,
bicarbonate, potassium or sodium acetate, ammonia, and Scott’s Tap water substitute (TWS).
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46. Autotechnicon makes use of 12 individual processing steps, with ten 1-liter capacity glass
beakers & 2 thermostatically controlled wax baths with a safety device cut-out switch to
protect the wax against over-heating. Autotechnicon is an automatic tissue processing
machine that fixes, dehydrates, clears & infiltrates tissues.
47. Major steps in the routine H&E staining includes: deparaffinization, hydration, nuclear
staining, differentiation, bluing, counterstaining, dehydration and clearing.
48. Ascending degrees of alcohol promotes dehydration, prior to exposure of tissues to xylol and
paraffin. Descending grades of alcohol promotes hydration.
49. Direct staining method is the process by which sections are stained with simple aqueous or
alcoholic solutions of the dye
50. Indirect staining is the process whereby the action of the dye is intensified by adding
another agent (mordant) which serves as a link or bridge between the tissue and the dye, to
make the staining reaction possible. By itself, the dye may stain only weakly, if at all. The
mordant combines with the dye to form a colored “lake”, which in turn combines with the
tissue to form a “tissue-mordant-dye-complex” that is rendered insoluble in ordinary
aqueous and alcoholic solvents.
51. Metachromatic dye imparts a color to the tissue that is different from the color of the dye
itself.
52. Metachromatic dyes are basic dyes belonging to the thiazine and triphenylmethane groups.
This includes: methyl violet or crystal violet, cresyl blue, safranin, Bismarck brown, basic
fuchsin, methylene blue, thionine, toluidine blue as well as azures A, B and C.
53. Example of nuclear stains: hematoxylin, methylene blue, toluidine blue and neutral red.
54. Hematoxylin is a natural dye derived by extraction from the core or the heartwood of a
Mexican tree known as Hematoxylin campechianum.
55. Hematoxylin itself is not a stain. The active coloring agent is hematin, which is formed by the
oxidation of hematoxylin, a process known as ripening. This is usually accomplished by
exposing the substance to air and sunlight, thereby oxidizing hematoxylin (natural ripening).
Such a process is slow and takes as long as 3-4 months, but it can be accelerated by adding
strong oxiding agents such as hydrogen peroxide, mercuric oxide, potassium permanganate,
sodium perborate or sodium iodate which converts hematoxylin to hematin almost
instantaneously be chemical oxidation (artificial ripening), so that staining solution is ready
for use immediately after the preparation.
56. Mordants for hematoxylin
▪ Aluminum (alum/potassium aluminum sulfate) = Ehrlich, Harris, Cole, Mayer
▪ Iron = Weigert (ferric ammonium chloride) and Heidenhain (ferric ammonium sulfate)
▪ Phosphotungstic acid = PTAH
▪ Chromium
57. In Routine H & E staining, tissues are differentiated in 1% acid alcohol after primary staining
with hematoxylin. 1% acid alcohol is formed by combination of 1mL of concentrated
hydrochloric acid to 99mL of 80% ethanol. Acid differentiation is done for approximately 10-
30 seconds, controlling microscopically until only the nuclei are stained. H&E staining is an
example of a regressive staining method.
58. Staining result of tissue components or types using routine H and E staining technique:
• Nuclei = blue to blue black
• Karyosome = dark blue
• Cytoplasm, proteins in edema fluid = pale pink
• RBCs, eosinophilic granules, keratin = bright orange red
• Basophil cytoplasm, plasma cells and osteoblast = purplish pink
• Cartilage = pink or light blue to dark blue, depending on the type of stain used
• Calcium and calcified bone = purplish blue
• Decalcified bone matrix, collagen and osteoid = pink
• Muscle fibers = deep pink
59. Weigert’s solution and Heidenhain’s solution are iron hematoxylin solutions employed for
routine work in the laboratory. Weigert’s solution uses ferric ammonium chloride and
Heidenhain’s solution uses ferric ammonium sulfate as mordant.
60. EA50 is made up of the following:
• Bismarck brown
• Eosin-Yellow
• Light Green SF yellow solution
• Phosphotungstic acid
• Lithium carbonate

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61. Methylene blue is a valuable stain for plasma cells and may also be employed in cytological
examinations of fresh sputum for malignant cells, as a bacterial stain for evaluation and
differentiation of bacterial organisms, for diphtheria and for vital staining of the nervous
system. ~p151, Histopathologic Techniques by Bruce-Gregorios
62. Methyl green-pyronin can be used to stain: DNA & RNA
63. Silver stains are used to stain reticulin fibers, calcium deposits, Legionella pneumophila and
spirochetes.
64. Collagen may be differentially stained by: (1) van Gieson’s stain; (2) Masson’s trichrome
stain; (3) Malory’s aniline blue stain; (4) azocarmine stain; (5) Krajian’s aniline blue stain.
65. Von Kossa’s silver nitrate method for calcium demonstration stain calcium salts black.
66. Elastic fibers are stained dark brown by orcein. –p209, Histopathologic Technique
67. Using Masson Fontana Technique for Melanin and argentaffin cells, melanin pigments and
argentaffin cell granules would stain BLACK and nuclei of cells will stain red.
68. Iodine is the oldest stain used for amyloid, starch and glycogen. It can also be used for the
removal of mercuric precipitates that forms during fixation.
69. PAS-positive substances stain red or magenta red.
70. For amyloid demonstration, the following methods may be used:
Gram’s iodine stain; Congo red method;
Metachromatic staining (methyl violet-crystal violet method) Induced fluorescence staining with
thioflavine
71. Glycogen is demonstrated using PAS (periodic acid schiff), Best Carmine and Langhan’s
iodine method.
72. Neutral red is a basic dye recommended for observing cell granules and vacuoles of
phagocytic cells.
73. For broken slides:
• mount the broken slide into another slide that has been moistened with xylene, with a
drop of mountant (either Permount or Clarite)
• section can be transferred to another slide if still intact
74. Specimens for exfoliative cytology include:
• Cervicovaginal smear, Nipple discharge,
• Gastric or bronchial secretions, Sputum,
• Pleural and peritoneal fluids, Urine sediment
• Cerebrospinal fluid
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75. Mayer’s egg albumin is an adhesive composed of equal parts of egg white and glycerol, with
the addition of thymol crystals. Thymol crystals are added to Mayer’s egg albumin to prevent
growth of fungus in the solution.
76. To avoid distortion of the image, the refractive index of the mountant should be as near as
possible to that of the glass, which is 1.518.
77. Kronig cement is a ringing media – it is used to seal the margins of the coverslip to prevent
escape of fluid and evaporation of mountant, as well as to immobilize the coverslip and to
prevent sticking of slides upon storage. It is composed of 2 parts paraffin wax, mixed with 4-
9 parts of powdered colophonium resin, heated and filtered.
78. In the Old Manner of Reporting Pap smear:
• Class I is characterized by the absence of atypical or abnormal cells
• Class II is described by the presence of atypical cytologic picture but no evidence of
malignancy
• Class III is characterized by a cytological picture suggestive but not conclusive of
malignancy
• Class IV is characterized by a cytological picture that is strongly suggestive of
malignancy
• Class V is characterized by a cytologic picture conclusive of malignancy.
79. Apoptosis refers to programmed cell death.
80. Pyknosis refers to condensation of chromatin material.
81. Karyorrhexis refers to breaking down of nuclear components or segmentation of nucleus.
82. Karyolysis refers to dissolution of the nuclear structures.
83. Mature superficial cells are polygonal squamous cells that measures 45 to 50μm in diameter
and are usually identified by the presence of pale, pink-staining cytoplasm and dark pyknotic
nuclei, which is less than 6μ in diameter
84. Parabasal cells are round to oval cells with small, dense, basophilic cytoplasm, and a total
cell diameter of 15 to 30μm. They are smaller than intermediate cells, and have a larger
vesicular nucleus; normally found from two weeks of age to puberty, after childbirth, with
abortions and after menopause
85. Intermediate cells are medium-sized, polyhedral or elongated cells with basophilic,
vacuolated cytoplasm
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86. Transitional epithelial cells originate from the lining of the renal pelvis, calyces, ureters, and • Dysplasia = regressive alteration in adult cells manifested by variation in size, shape and
bladder, and from the upper portion of the male urethra. Squamous epithelial cells originate orientation, associated with chronic inflammation and protracted irritation; reversible
from the linings of the vagina and female urethra and the lower portion of the male urethra. and do not inevitably lead to tumor formation
87. Cardinal Signs of Inflammation: • Anaplasia = Dedifferentiation; marked regressive change in adult cells towards a more
• Rubor (redness) = due to arteriolar and capillary dilatation with increased rate of blood primitive or embryonic cell type, usually utilized as a criterion towards malignancy;
flow towards the site of the injury and concentration or packing of the red cells in the irreversible
capillaries causing increased viscosity and consequent slowing of the blood flow • Neoplasia = continuous abnormal proliferation of cells without control, serving no useful
• Tumor (swelling) = due to increased capillary permeability, allowing the extravasation purpose or function, usually accompanied by increase in size and pigmentation, mitosis,
of blood fluid, associated with increased hydrostatic pressure within the dilated number, metaplastic and anaplastic changes of the cell
arterioles and capillaries, causing localized edema (tumor) accompanied by escape of
blood cells into the injured area, increased concentration of plasma proteins and
accumulation of finely particulate cellular debris caused by destruction of tissue cells
and bacteria.
• Calor (heat) = due to transfer of internal heat to the surface or site of injury brought
about by the increased blood content
• Dolor (pain) = due to the pressure upon the sensory nerve by the exudate or tumor
• Functio Laesa (loss of function) = may be due to pain interference with nerve supply and
to destruction of the functioning units of the tissue
88. Tissue Changes
• Aplasia = incomplete or defective development of a tissue or organ, represented only by
a mass of fatty or fibrous tissue, bearing no resemblance to adult structure
• Agenesia = refers to complete nonappearance of an organ
• Hypoplasia = refers to the failure of an organ to reach or achieve its full mature or adult
size due to incomplete development
• Atresia = failure of an organ to form an opening
• Atrophy = refers to an acquired decrease in the size of a normally developed or mature
tissue or organ, resulting from reduction in cell size or decrease in total number of cells
or both
• Hypertrophy = refers to an increase in size of tissues or organs due to increase in the
size of the individual cells
• Hyperplasia = refers to increase in size of an organ or tissue due to increase in the
number of cells resulting from growth of new cells
• Metaplasia = reversible transformation from one type of adult cell into another

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MEDTECH LAWS AND ETHICS ▪ Section 27: Foreign Reciprocity

▪ Section 28: Roster of Medical Technologist
▪ Section 29: Penal Provision
1. RA5527 – Philippine Medical Technology Act of 1969
▪ Section 30: Separability Clause
▪ Section 1: Title
▪ Section 31: Repealing Clause
▪ Section 2: Definition of Terms
▪ Section 32: Effectivity
▪ Section 3: Council of Medical Technology Education, its Composition
2. Board of Medical Technology, according to RA 5527 is composed of a chairman, which is a
▪ Section 4: Compensation & traveling expenses of the Council members
pathologist and two medical technologists as members.
▪ Section 5: Functions of the Council of Medical Technology Education
3. In order to pass the examination, a candidate must obtain a general average of at least 75%
▪ Section 6: Minimum required course
in the written test, with no rating below 50% in any of the major subjects: provided that, that
▪ Section 7: Medical Technology Board
candidate has not failed in at least 60% of the subjects computed according to their relative
▪ Section 8: Qualification of Examiners
weights.
▪ Section 9: Executive Officer of the Board
4. The medtech board shall likewise issue a certificate of registration as a medical laboratory
▪ Section 10: Compensation of Members of Members of the Board Examiners for Medical
technician provided that he/she has failed to pass the examination for medical technology
Technology
but had obtained a general obtained a general rating of at least 70%.~section 21 (Issuance of
▪ Section 11: Functions and Duties of the Board
Certificate of Registration) of RA 5527
▪ Section 12: Removal of Board Members
5. Section 5 of RA 5527 states the functions of the council of medical technology education:
▪ Section 13: Accreditation of Schools of Medical technology and of training Laboratories
• to recommend the minimum required curriculum for the course of medtech
▪ Section 14: Inhibition against the Practice of Medical Technology
• to determine and prescribe the number of students allowed to take medical technology
▪ Section 15: Examination
course each school
▪ Section 16: Qualification for Examination
• to approve medical technology schools, taking into account the student-instructor-ratio
▪ Section 17: Scope of Examination
and availability of facilities
▪ Section 18: Report of Rating
• to require all medtech schools to submit an annual report
▪ Section 19: Rating in the Examination
• to inspect when necessary, the different medtech schools
▪ Section 20: Oath Taking
• to certify for admission into an undergraduate internship students who have
▪ Section 21: Issuance of Certificate of Registration
satisfactorily completed three years of medtech course
▪ Section 22: Fees
• formulate and recommend approval of refresher course for applicants who shall have
▪ Section 23: Refusal to Issue Certificate
failed the Board Examination for the third time
▪ Section 24: Administrative Investigation
• to promulgate the prescribe and enforce necessary rules and regulations for proper
▪ Section 25: Appeal
implementation of functions
▪ Section 26: Reinstatement, Reissue and Replacement of Certificate

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6. Section 11 of RA 5527 state the functions and duties of the Board of Medical Technology
▪ administer the provisions of RA5527
▪ administer oaths
▪ issue, suspend, and revoke certificate of registration
▪ look into conditions affecting the practice of medical technology in the Philippines
▪ investigate such violators of RA5527
▪ draft rules and regulations to carry out provision of RA5527
▪ to determine the adequacy of technical staff of all clinical laboratories and blood banks
▪ to prescribe the qualifications and training of medical technologist as to special field of
the profession
▪ to classify and prescribe the qualifications and training of technical staff of clinical
laboratories as to: medical technology, medical technologist, senior medical
technologist, medical technologist and medical laboratory technician
7. According to RA 8981 (PRC Modernization Act of 2000), the chairperson or commissioner
should be:
▪ at least 40 years of age; holds a valid certificate of registration/professional license and
a valid professional identification card; or a valid certificate of competency issued by the
commission or a valid professional license issued by any government agency
▪ familiar with the principles and methods of professional regulation and/licensing
▪ had at least 5 years of executive or management experience: provided that one of the
commissioners must be a past chairperson/member of a professional regulatory board
8. Medtech-related Laws
• RA 5527 = Philippine Medical Technology Act of 1969
• RA 8981 = The PRC Modernization Act of 2000
• RA 1517 = Blood Banking Law of the Philippines
• RA 7719 = National Voluntary Blood Services Act
• RA 8504 = Philippine HIV/AIDS Law
• RA 4688 = Clinical Laboratory Law
• RA 6425 = Dangerous Drug Act of 1972
• RA 9165 = Dangerous Drug Act of 2002
• RA 9288 = Newborn Screening Act of 2004
• RA 7170 = Organ Donation Act 1991
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9. PD 1534 amended RA 5527 on June 11, 1978. It amended sections 3, 8 and 13 of RA 5527.
10. Clinical laboratories shall be inspected every two years or as necessary. Blood banks are
inspected yearly
11. The following are included in the Newborn Screening procedure: phenylketonuria,
galactosemia, G6PD deficiency, congenital hypothyroidism or cretinism, and congenital
adrenal hyperplasia. Latest addition to the group is the maple syrup urine disease (MSUD)
12. PRC resolution no. 323 includes the policies on admission of foreigners to the licensure
examinations and the practice of their profession.
13. Executive order no. 266 - Institutionalization of the Continuing Professional Education (CPE)
programs of the various Professional Regulatory Boards (PRBs) under the supervision of the
Professional Regulation Commission
14. 60 CPE points are required for the renewal of medical technology license. Renewal of PRC
license is done every 3 years
15. Republic Act No. 10912, otherwise known as the “Continuing Professional Development
(CPD) Act of 2016”, is an act which requires CPD as the mandatory requirement for the
renewal of Professional Identification Card. Based on Board Resolution No. 07, S.2018 of
Medical Technology, medical technologists are required to have 45 CPD Credit units while
medical technicians are required to have 30 CPD Credit units.
16. PAMET HYMN
Music by Francis Jerota Pefanco
Lyrics by Hector G. Gayares, Jr.
17. PANUNUMPA NG PROPESYONAL
Ako, si (pangalan), ng (pook na sinilangan, bayan/lungsod, probinsya) ay
taimtim na nanunumpa na itataguyod ko at ipagtatanggol ang Saligang Batas ng Pilipinas,
na ako ay tunay na mananalig at tatalima rito; na susundin ko ang mga batas, mga utos na
legal, at mga atas na ipinahayag ng mga sadyang itinakdang may
kapangyarihan ng Republika ng Pilipinas; at kusa kong babalikatin ang pananagutang ito,
na walang ano mang pasubali o hangaring umiwas.
Taimtim pa rin akong manunumpa na sa lahat ng panahon at pook na kinaroroonan ay
mahigpit akong manghahawakan sa mga etikal at tuntuning propesyonal ng mga Medical
Technologist sa Pilipinas, at marapat kong gagampanan ng buong husay sa
abot ng aking makakaya ang mga tungkulin at pananagutang iniatang sa isang
itinakdang Medical Technologist. Kasihan Nawa ako ng Diyos.
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18. CODE OF ETHICS
As I enter into the practice of Medical Technology, I shall accept the responsibilities inherent
to being a professional.
I shall uphold the law and shall not engage in illegal work nor cooperate with anyone so
engaged;
I shall avoid associating or being identified with any enterprise of questionable character;
I shall work in strict spirit of fairness to employer, clients, contractors and employees and in
a spirit of personal helpfulness and fraternity toward other members of the profession;
I shall use only honorable means of competition for professional employment or services
and shall refrain from unfairly injuring, directly or indirectly, the professional reputation,
projects or business of a fellow medical technologist;
I shall accept employment from more than one employer only when there is no conflict of
interest;
I shall perform professional work in a manner that merits full confidence and trust carried
out with absolute reliability, accuracy, fairness and honesty;
I shall review the professional work of other medical technologists, when requested, fairly
and in confidence whether they are employees, authors of proposals or grants or contracts,
authors of technical papers or other publications or involved litigation; I shall advance the
profession by exchanging general information and experience with fellow medical
technologists and other professionals and be contributing to the work of professional
organizations;
I shall restrict my praises, criticisms, views and opinions within constructive limits and shall
not use the knowledge I know for selfish ends;
I shall treat any information I acquired about individuals in the course of my work as strictly
confidential, and may be divulged only to authorized persons or entities or with consent of
the individual when necessary;
I shall report any infarctions of these principles of professional conduct to the authorities
responsible of enforcement of applicable laws or regulations or to the Ethics Committee of
the Philippine Association of Medical Technologist as may be appropriate.
To these principles, I hereby subscribe and pledge to conduct myself at all times in a manner
befitting the dignity of my profession.
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COVID-19: FACT SHEET
(from CDC,WHO, webMD, Roche, etc. as of Jan 8, 2021)
1. Coronavirus Disease 2019 (COVID-19) caused by a coronavirus, SARS-CoV-2.
2. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is an enveloped, single-
stranded RNA virus of the family Coronaviridae. Coronaviruses share structural similarities
and are composed of 16 nonstructural proteins and 4 structural proteins: spike (S), envelope
(E), membrane (M), and nucleocapsid (N).
3. The virus accesses host cells via the angiotensin-converting enzyme 2 (ACE2) receptor, which
is most abundant in the lungs
4. WHO first learned of this new virus on 31 December 2019, following a report of a cluster of
cases of ‘viral pneumonia’ in Wuhan, People’s Republic of China.
5. Experts say SARS-CoV-2 originated in bats. SARS-CoV-2 made the jump to humans at one of
Wuhan’s open-air “wet markets.” They’re where customers buy fresh meat and fish, including
animals that are killed on the spot.
6. The Wuhan market didn’t sell bats at the time of the outbreak. That’s why early suspicion also
fell on pangolins, also called scaly anteaters, which are sold illegally in some markets in China.
Some coronaviruses that infect pangolins are similar to SARS-CoV-2.
7. COVID-19 is spread through respiratory droplets when an infected person coughs, sneezes or
speaks. People can also be infected by touching a contaminated surface and then their eyes,
mouth or nose. You can become infected by coming into close contact (about 6 feet or two arm
lengths) with a person who has COVID-19.
8. The most common symptoms of COVID-19 are fever, dry cough, fatigue, loss of smell and taste,
shortness of breath
9. People aged 60 years and over, and those with underlying medical problems like high blood
pressure, heart and lung problems, diabetes, obesity or cancer, are at higher risk of developing
serious illness.
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10. Viral (nucleic acid or antigen) tests check samples from the respiratory system (such as nasal
or oral swabs or saliva) to determine whether an infection with SARS-CoV-2, the virus that
causes COVID-19, is present. Viral tests are recommended to diagnose acute infection of both
symptomatic and asymptomatic individuals, to guide contact tracing, treatment options, and
isolation requirements Some tests are point-of-care tests, meaning results may be available at
the testing site in less than an hour. Other tests must be sent to a laboratory, a process that
may take at least 1-2 days
11. Rapid diagnostic tests (RDT) detect the presence of viral proteins (antigens) expressed by the
COVID-19 virus in a sample from the respiratory tract of a person.
12. Most Ag-RDTs for COVID-19 use a sandwich immunodetection method employing a simple-
to-use lateral flow test format commonly employed for HIV, malaria and influenza testing. Ag-
RDTs are usually comprised of a plastic cassette with sample and buffer wells, a nitrocellulose
matrix strip, with a test line with bound antibody specific for conjugated target antigen-
antibody complexes and a control line with bound antibody specific for conjugated-antibody.
In the case of SARS-CoV-2 RDTs the target analyte is often the virus’ nucleocapsid protein,
preferred because of its relative abundance.
13. Real-Time chemistries allow for the detection of PCR amplification during the early phases of
the reaction. Measuring the kinetics of the reaction in the early phases of PCR provides a
distinct advantage over traditional PCR detection. Traditional methods use Agarose gels for
detection of PCR amplification at the final phase or end-point of the PCR reaction
14. RT PCR cannot be used to detect past infections.
15.
\ CDC does not currently recommend using antibody testing as the sole basis for diagnosis of
acute infection.
16. Additional Information About Covid-19
▪ Studies show hydroxychloroquine does not have clinical benefits in treating COVID-19
▪ Water or swimming does not transmit the COVID-19 virus
▪ Drinking alcohol does not protect you against COVID-19 and can be dangerous. Adding
pepper to your soup or other meals DOES NOT prevent or cure COVID-19. Eating garlic
does NOT prevent COVID-19
▪ The COVID-19 virus CANNOT be spread through mosquito bites
▪ COVID-19 is NOT transmitted through houseflies
▪ Spraying and introducing bleach or another disinfectant into your body WILL NOT protect
you against COVID-19 and can be dangerous
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▪ Exposing yourself to the sun or temperatures higher than 25°C DOES NOT protect you
from COVID-19
▪ The COVID-19 virus can spread in hot and humid climates. Cold weather and snow
CANNOT kill the COVID-19 virus.
▪ Taking a hot bath does not prevent COVID-19. Hand dryers are NOT effective in killing the
COVID-19 virus. Ultra-violet (UV) lamps should NOT be used to disinfect hands or other
areas of your skin
▪ Rinsing your nose with saline does NOT prevent COVID-19
▪ Vaccines against pneumonia DO NOT protect against the COVID-19 virus
▪ Current vaccines being developed to date (by Pfizer and Moderna) contains mRNA
(messenger RNA). These mRNAs give instruction to the body’s cells to make harmless
proteins known as spike proteins, which is found on the surface of SARS-CoV-2. This
protein piece is displayed on the cell surface, triggering our immune system to make
antibodies against it.
▪ Some vaccines being currently developed to date (by AstraZeneca) contains weakened
version of a common cold virus, the adenovirus taken from chimpanzees. The adenovirus
is genetically altered making them unable to reproduce. The vaccine is combined with
genes of the spike protein to trigger production of antibodies against it. Altered adenovirus t
spike
-
▪ People of all ages can be infected by the COVID-19 virus protein
▪ Antibiotics CANNOT prevent or treat COVID-19. Antibiotics work only against bacteria,
not viruses. However, if you are hospitalized for COVID-19, you may receive antibiotics
because bacterial co-infection is possible.
▪ Catching COVID-19 DOES NOT mean you will have it for life
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